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<article doi="10.1038/35044114" abstract="Autophagy is a dynamic membrane phenomenon for bulk protein degradation in the lysosome/vacuole. Apg8/Aut7 is an essential factor for autophagy in yeast. We previously found that the carboxy-terminal arginine of nascent Apg8 is removed by Apg4/Aut2 protease, leaving a glycine residue at the C terminus. Apg8 is then converted to a form (Apg8-X) that is tightly bound to the membrane. Here we report a new mode of protein lipidation. Apg8 is covalently conjugated to phosphatidylethanolamine through an amide bond between the C-terminal glycine and the amino group of phosphatidylethanolamine. This lipidation is mediated by a ubiquitination-like system. Apg8 is a ubiquitin-like protein that is activated by an E1 protein, Apg7 (refs 7, 8), and is transferred subsequently to the E2 enzymes Apg3/Aut1 (ref. 9). Apg7 activates two different ubiquitin-like proteins, Apg12 (ref. 10) and Apg8, and assigns them to specific E2 enzymes, Apg10 (ref. 11) and Apg3, respectively. These reactions are necessary for the formation of Apg8-phosphatidylethanolamine. This lipidation has an essential role in membrane dynamics during autophagy."><fig id="2632"><title></title><label>figf1</label><graphic href="https://api.sourcedata.io/file.php?figure_id=2632"/><sd-panel panel_id="9126"><strong>a</strong><span>, <sd-tag id="sdTag1" source="sdapi" category="assay" entity_type="" role="" text="Phase partitioning" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">Phase partitioning</sd-tag> of <sd-tag id="sdTag2" source="sdapi" category="entity" entity_type="protein" role="assayed" text="Apg8" ext_ids="P38182" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="ATG8" ext_tax_ids="559292" ext_tax_names="Saccharomyces cerevisiae S288C" ext_urls="https://www.uniprot.org/uniprot/">Apg8</sd-tag>. Wild-type and Δ</span><em><sd-tag id="sdTag3" source="sdapi" category="entity" entity_type="gene" role="intervention" text="apg4" ext_ids="855498" norm_text="" ext_dbs="NCBI gene" in_caption="True" ext_names="ATG4" ext_tax_ids="559292" ext_tax_names="Saccharomyces cerevisiae S288C" ext_urls="http://www.ncbi.nlm.nih.gov/gene/">apg4</sd-tag></em>Δ<em><sd-tag id="sdTag4" source="sdapi" category="entity" entity_type="gene" role="intervention" text="apg8" ext_ids="852200" norm_text="" ext_dbs="NCBI gene" in_caption="True" ext_names="ATG8" ext_tax_ids="559292" ext_tax_names="Saccharomyces cerevisiae S288C" ext_urls="http://www.ncbi.nlm.nih.gov/gene/">apg8</sd-tag></em> cells producing Apg8FG and Δ<em><sd-tag id="sdTag6" source="sdapi" category="entity" entity_type="gene" role="intervention" text="apg7" ext_ids="856576" norm_text="" ext_dbs="NCBI gene" in_caption="True" ext_names="ATG7" ext_tax_ids="559292" ext_tax_names="Saccharomyces cerevisiae S288C" ext_urls="http://www.ncbi.nlm.nih.gov/gene/">apg7</sd-tag></em> cells were subjected to phase partitioning (see Methods).<graphic href="https://api.sourcedata.io/file.php?panel_id=9126"/></sd-panel><sd-panel panel_id="9127"><strong>b</strong>, MALDI mass spectrum of <sd-tag id="sdTag9" source="sdapi" category="entity" entity_type="molecule" role="reporter" text="His6" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">His<sub>6</sub></sd-tag>-<sd-tag id="sdTag13" source="sdapi" category="entity" entity_type="protein" role="assayed" text="Apg8" ext_ids="P38182" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="ATG8" ext_tax_ids="559292" ext_tax_names="Saccharomyces cerevisiae S288C" ext_urls="https://www.uniprot.org/uniprot/">Apg8</sd-tag>. Purified <sd-tag id="sdTag10" source="sdapi" category="entity" entity_type="molecule" role="reporter" text="His6" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">His<sub>6</sub></sd-tag>-<sd-tag id="sdTag14" source="sdapi" category="entity" entity_type="protein" role="assayed" text="Apg8" ext_ids="P38182" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="ATG8" ext_tax_ids="559292" ext_tax_names="Saccharomyces cerevisiae S288C" ext_urls="https://www.uniprot.org/uniprot/">Apg8</sd-tag> (predominantly <span><sd-tag id="sdTag154" source="sdapi" category="entity" entity_type="protein" role="assayed" text="Apg8" ext_ids="P38182" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="" ext_tax_ids="559292" ext_tax_names="Saccharomyces cerevisiae S288C" ext_urls="https://www.uniprot.org/uniprot/">Apg8</sd-tag>FG</span>-X and a small amount of Apg8FG<span>) was extracted with 0.4% <sd-tag id="sdTag11" source="sdapi" category="entity" entity_type="molecule" role="component" text="β-octylglycoside" ext_ids="CHEBI:41128" norm_text="" ext_dbs="ChEBI" in_caption="True" ext_names="B-OCTYLGLUCOSIDE" ext_tax_ids="" ext_tax_names="" ext_urls="http://www.ebi.ac.uk/chebi/searchId.do?chebiId=">β-octylglycoside</sd-tag> from a polyacrylamide gel and desalted with</span> ZipTip<sub>C4</sub>(Millipore) before mass measurement. The principal MH+ ion signal at <em>m</em>/<em>z</em> 15,003 is shifted in mass by about +635 from the MH+ value corresponding to His<sub>6</sub>-Apg8FG.<graphic href="https://api.sourcedata.io/file.php?panel_id=9127"/></sd-panel><sd-panel panel_id="9129"><strong>c</strong><span>, <sd-tag id="sdTag18" source="sdapi" category="assay" entity_type="" role="" text="MS/MS" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">MS/MS</sd-tag> spectra of the C-terminal peptides of</span> <span><sd-tag id="sdTag156" source="sdapi" category="entity" entity_type="protein" role="assayed" text="Apg8" ext_ids="P38182" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="" ext_tax_ids="559292" ext_tax_names="Saccharomyces cerevisiae S288C" ext_urls="https://www.uniprot.org/uniprot/">Apg8</sd-tag>FG</span>-X (top) and Apg8FG (bottom). C-terminal fragments were generated from Apg8FG-X and Apg8FG <span>(see Methods). Each fragment that was observed only as non-18O-labelled signals was subjected to <sd-tag id="sdTag24" source="sdapi" category="assay" entity_type="" role="" text="MS/MS" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">MS/MS</sd-tag>. The <sd-tag id="sdTag25" source="sdapi" category="assay" entity_type="" role="" text="MS/MS" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">MS/MS</sd-tag> spectra, deconvoluted with MaxEnt3, were interpreted by SeqMS software26. Arrows show the sequences from the N and C terminus based on y″</span><sub><em>m</em></sub> and b<sub><em>l</em></sub> ions, respectively (where <em>m</em> and <em>l</em> <span>denote positions counted from the C and N terminus) that were produced by cleavage of peptide bonds during <sd-tag id="sdTag26" source="sdapi" category="assay" entity_type="" role="" text="MS/MS" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">MS/MS</sd-tag>. Amino acids shown in three-letter code denote <sd-tag id="sdTag19" source="sdapi" category="entity" entity_type="molecule" role="assayed" text="immonium ions" ext_ids="CHEBI:35286" norm_text="" ext_dbs="ChEBI" in_caption="True" ext_names="iminium ion" ext_tax_ids="" ext_tax_names="" ext_urls="http://www.ebi.ac.uk/chebi/searchId.do?chebiId=">immonium ions</sd-tag>. The nomenclature of these ions is as in previous work28.</span><graphic href="https://api.sourcedata.io/file.php?panel_id=9129"/></sd-panel></fig><fig id="2633"><title></title><label>figf2</label><graphic href="https://api.sourcedata.io/file.php?figure_id=2633"/><sd-panel panel_id="9134"><strong>d</strong>, <sd-tag id="sdTag133" source="sdapi" category="entity" entity_type="molecule" role="component" text="Thioester" ext_ids="CHEBI:51277" norm_text="" ext_dbs="ChEBI" in_caption="True" ext_names="thioester" ext_tax_ids="" ext_tax_names="" ext_urls="http://www.ebi.ac.uk/chebi/searchId.do?chebiId=">Thioester</sd-tag> formation between <sd-tag id="sdTag134" source="sdapi" category="entity" entity_type="protein" role="assayed" text="Apg8" ext_ids="P38182" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="ATG8" ext_tax_ids="559292" ext_tax_names="Saccharomyces cerevisiae S288C" ext_urls="https://www.uniprot.org/uniprot/">Apg8</sd-tag> and <sd-tag id="sdTag135" source="sdapi" category="entity" entity_type="protein" role="assayed" text="Apg7" ext_ids="P38862" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="ATG7" ext_tax_ids="559292" ext_tax_names="Saccharomyces cerevisiae S288C" ext_urls="https://www.uniprot.org/uniprot/">Apg7</sd-tag>. Cell extracts of Δ<em><sd-tag id="sdTag136" source="sdapi" category="entity" entity_type="gene" role="intervention" text="apg7" ext_ids="856576" norm_text="" ext_dbs="NCBI gene" in_caption="True" ext_names="ATG7" ext_tax_ids="559292" ext_tax_names="Saccharomyces cerevisiae S288C" ext_urls="http://www.ncbi.nlm.nih.gov/gene/">apg7</sd-tag></em>Δ<em><sd-tag id="sdTag137" source="sdapi" category="entity" entity_type="gene" role="intervention" text="apg8" ext_ids="852200" norm_text="" ext_dbs="NCBI gene" in_caption="True" ext_names="ATG8" ext_tax_ids="559292" ext_tax_names="Saccharomyces cerevisiae S288C" ext_urls="http://www.ncbi.nlm.nih.gov/gene/">apg8</sd-tag></em> cells expressing <sd-tag id="sdTag138" source="sdapi" category="entity" entity_type="protein" role="assayed" text="Apg7" ext_ids="P38862" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="ATG7" ext_tax_ids="559292" ext_tax_names="Saccharomyces cerevisiae S288C" ext_urls="https://www.uniprot.org/uniprot/">Apg7</sd-tag>-<sd-tag id="sdTag139" source="sdapi" category="entity" entity_type="protein" role="reporter" text="Myc" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">Myc</sd-tag> and/or <sd-tag id="sdTag140" source="sdapi" category="entity" entity_type="protein" role="reporter" text="HA" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">HA</sd-tag>-<sd-tag id="sdTag141" source="sdapi" category="entity" entity_type="protein" role="assayed" text="Apg8" ext_ids="P38182" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="ATG8" ext_tax_ids="559292" ext_tax_names="Saccharomyces cerevisiae S288C" ext_urls="https://www.uniprot.org/uniprot/">Apg8</sd-tag> from 2µ plasmids were <sd-tag id="sdTag142" source="sdapi" category="assay" entity_type="" role="" text="immunoprecipitated" ext_ids="BAO_0002508" norm_text="" ext_dbs="BAO" in_caption="True" ext_names="immunoprecipitation" ext_tax_ids="" ext_tax_names="" ext_urls="https://bioportal.bioontology.org/ontologies/BAO/?p=classes&conceptid=http%3A%2F%2Fwww.bioassayontology.org%2Fbao%23">immunoprecipitated</sd-tag> with anti-<sd-tag id="sdTag143" source="sdapi" category="entity" entity_type="protein" role="reporter" text="HA" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">HA</sd-tag> antibody, and <sd-tag id="sdTag144" source="sdapi" category="assay" entity_type="" role="" text="immunoblotted" ext_ids="BAO_0002424" norm_text="" ext_dbs="BAO" in_caption="True" ext_names="western blot" ext_tax_ids="" ext_tax_names="" ext_urls="https://bioportal.bioontology.org/ontologies/BAO/?p=classes&conceptid=http%3A%2F%2Fwww.bioassayontology.org%2Fbao%23">immunoblotted</sd-tag> (IB) with anti-<sd-tag id="sdTag145" source="sdapi" category="entity" entity_type="protein" role="reporter" text="Myc" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">Myc</sd-tag> and anti-<sd-tag id="sdTag146" source="sdapi" category="entity" entity_type="protein" role="reporter" text="HA" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">HA</sd-tag> antibody. <sd-tag id="sdTag147" source="sdapi" category="entity" entity_type="molecule" role="intervention" text="DTT" ext_ids="CHEBI:18320" norm_text="" ext_dbs="ChEBI" in_caption="True" ext_names="1,4-dithiothreitol" ext_tax_ids="" ext_tax_names="" ext_urls="http://www.ebi.ac.uk/chebi/searchId.do?chebiId=">DTT</sd-tag>-, no <sd-tag id="sdTag148" source="sdapi" category="entity" entity_type="molecule" role="intervention" text="DTT" ext_ids="CHEBI:18320" norm_text="" ext_dbs="ChEBI" in_caption="True" ext_names="1,4-dithiothreitol" ext_tax_ids="" ext_tax_names="" ext_urls="http://www.ebi.ac.uk/chebi/searchId.do?chebiId=">DTT</sd-tag>; <sd-tag id="sdTag149" source="sdapi" category="entity" entity_type="molecule" role="intervention" text="DTT" ext_ids="CHEBI:18320" norm_text="" ext_dbs="ChEBI" in_caption="True" ext_names="1,4-dithiothreitol" ext_tax_ids="" ext_tax_names="" ext_urls="http://www.ebi.ac.uk/chebi/searchId.do?chebiId=">DTT</sd-tag>+, 100 mM <sd-tag id="sdTag150" source="sdapi" category="entity" entity_type="molecule" role="intervention" text="DTT" ext_ids="CHEBI:18320" norm_text="" ext_dbs="ChEBI" in_caption="True" ext_names="1,4-dithiothreitol" ext_tax_ids="" ext_tax_names="" ext_urls="http://www.ebi.ac.uk/chebi/searchId.do?chebiId=">DTT</sd-tag>; asterisk, <sd-tag id="sdTag151" source="sdapi" category="entity" entity_type="protein" role="assayed" text="Apg8" ext_ids="P38182" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="ATG8" ext_tax_ids="559292" ext_tax_names="Saccharomyces cerevisiae S288C" ext_urls="https://www.uniprot.org/uniprot/">Apg8</sd-tag>-<sd-tag id="sdTag152" source="sdapi" category="entity" entity_type="protein" role="assayed" text="Apg7" ext_ids="P38862" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="ATG7" ext_tax_ids="559292" ext_tax_names="Saccharomyces cerevisiae S288C" ext_urls="https://www.uniprot.org/uniprot/">Apg7</sd-tag> conjugate.<graphic href="https://api.sourcedata.io/file.php?panel_id=9134"/></sd-panel><sd-panel panel_id="9130"><p><strong>b</strong>, Interaction between <sd-tag id="sdTag33" source="sdapi" category="entity" entity_type="protein" role="assayed" text="Apg8" ext_ids="P38182" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="ATG8" ext_tax_ids="559292" ext_tax_names="Saccharomyces cerevisiae S288C" ext_urls="https://www.uniprot.org/uniprot/">Apg8</sd-tag> and <sd-tag id="sdTag27" source="sdapi" category="entity" entity_type="protein" role="assayed" text="Apg7" ext_ids="P38862" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="ATG7" ext_tax_ids="559292" ext_tax_names="Saccharomyces cerevisiae S288C" ext_urls="https://www.uniprot.org/uniprot/">Apg7</sd-tag>. Δ<em><sd-tag id="sdTag34" source="sdapi" category="entity" entity_type="gene" role="intervention" text="apg7" ext_ids="856576" norm_text="" ext_dbs="NCBI gene" in_caption="True" ext_names="ATG7" ext_tax_ids="559292" ext_tax_names="Saccharomyces cerevisiae S288C" ext_urls="http://www.ncbi.nlm.nih.gov/gene/">apg7</sd-tag></em>Δ<em><sd-tag id="sdTag35" source="sdapi" category="entity" entity_type="gene" role="intervention" text="apg8" ext_ids="852200" norm_text="" ext_dbs="NCBI gene" in_caption="True" ext_names="ATG8" ext_tax_ids="559292" ext_tax_names="Saccharomyces cerevisiae S288C" ext_urls="http://www.ncbi.nlm.nih.gov/gene/">apg8</sd-tag></em> cells were transformed with <sd-tag id="sdTag43" source="sdapi" category="entity" entity_type="gene" role="component" text="APG7" ext_ids="856576" norm_text="" ext_dbs="NCBI gene" in_caption="True" ext_names="ATG7" ext_tax_ids="559292" ext_tax_names="Saccharomyces cerevisiae S288C" ext_urls="http://www.ncbi.nlm.nih.gov/gene/">APG7</sd-tag>-<sd-tag id="sdTag29" source="sdapi" category="entity" entity_type="protein" role="reporter" text="Myc" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">Myc</sd-tag> and/or <sd-tag id="sdTag44" source="sdapi" category="entity" entity_type="gene" role="component" text="APG8" ext_ids="852200" norm_text="" ext_dbs="NCBI gene" in_caption="True" ext_names="ATG8" ext_tax_ids="559292" ext_tax_names="Saccharomyces cerevisiae S288C" ext_urls="http://www.ncbi.nlm.nih.gov/gene/">APG8</sd-tag> on 2µ plasmids. Cell lysates were <sd-tag id="sdTag31" source="sdapi" category="assay" entity_type="" role="" text="immunoprecipitated" ext_ids="BAO_0002508" norm_text="" ext_dbs="BAO" in_caption="True" ext_names="immunoprecipitation" ext_tax_ids="" ext_tax_names="" ext_urls="https://bioportal.bioontology.org/ontologies/BAO/?p=classes&conceptid=http%3A%2F%2Fwww.bioassayontology.org%2Fbao%23">immunoprecipitated</sd-tag> with anti-<sd-tag id="sdTag36" source="sdapi" category="entity" entity_type="protein" role="reporter" text="Myc" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">Myc</sd-tag> antibody, and <sd-tag id="sdTag32" source="sdapi" category="assay" entity_type="" role="" text="immunoblotted" ext_ids="BAO_0002424" norm_text="" ext_dbs="BAO" in_caption="True" ext_names="western blot" ext_tax_ids="" ext_tax_names="" ext_urls="https://bioportal.bioontology.org/ontologies/BAO/?p=classes&conceptid=http%3A%2F%2Fwww.bioassayontology.org%2Fbao%23">immunoblotted</sd-tag> with anti-<sd-tag id="sdTag37" source="sdapi" category="entity" entity_type="protein" role="reporter" text="Myc" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">Myc</sd-tag> or anti-<sd-tag id="sdTag38" source="sdapi" category="entity" entity_type="protein" role="assayed" text="Apg8" ext_ids="P38182" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="ATG8" ext_tax_ids="559292" ext_tax_names="Saccharomyces cerevisiae S288C" ext_urls="https://www.uniprot.org/uniprot/">Apg8</sd-tag> antibody. <sd-tag id="sdTag39" source="sdapi" category="entity" entity_type="protein" role="assayed" text="Apg8" ext_ids="P38182" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="ATG8" ext_tax_ids="559292" ext_tax_names="Saccharomyces cerevisiae S288C" ext_urls="https://www.uniprot.org/uniprot/">Apg8</sd-tag> co-<sd-tag id="sdTag40" source="sdapi" category="assay" entity_type="" role="" text="immunoprecipitated" ext_ids="BAO_0002508" norm_text="" ext_dbs="BAO" in_caption="True" ext_names="immunoprecipitation" ext_tax_ids="" ext_tax_names="" ext_urls="https://bioportal.bioontology.org/ontologies/BAO/?p=classes&conceptid=http%3A%2F%2Fwww.bioassayontology.org%2Fbao%23">immunoprecipitated</sd-tag> with <sd-tag id="sdTag41" source="sdapi" category="entity" entity_type="protein" role="assayed" text="Apg7" ext_ids="P38862" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="ATG7" ext_tax_ids="559292" ext_tax_names="Saccharomyces cerevisiae S288C" ext_urls="https://www.uniprot.org/uniprot/">Apg7</sd-tag>-<sd-tag id="sdTag42" source="sdapi" category="entity" entity_type="protein" role="reporter" text="Myc" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">Myc</sd-tag>.</p><graphic href="https://api.sourcedata.io/file.php?panel_id=9130"/></sd-panel><sd-panel panel_id="9132"><p><strong>c</strong>, Effect of <sd-tag id="sdTag45" source="sdapi" category="entity" entity_type="protein" role="assayed" text="Apg8" ext_ids="P38182" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="ATG8" ext_tax_ids="559292" ext_tax_names="Saccharomyces cerevisiae S288C" ext_urls="https://www.uniprot.org/uniprot/">Apg8</sd-tag> C-terminal processing on the <sd-tag id="sdTag46" source="sdapi" category="entity" entity_type="protein" role="assayed" text="Apg8" ext_ids="P38182" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="ATG8" ext_tax_ids="559292" ext_tax_names="Saccharomyces cerevisiae S288C" ext_urls="https://www.uniprot.org/uniprot/">Apg8</sd-tag>-<sd-tag id="sdTag47" source="sdapi" category="entity" entity_type="protein" role="assayed" text="Apg7" ext_ids="P38862" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="ATG7" ext_tax_ids="559292" ext_tax_names="Saccharomyces cerevisiae S288C" ext_urls="https://www.uniprot.org/uniprot/">Apg7</sd-tag> interaction. Δ<em><sd-tag id="sdTag48" source="sdapi" category="entity" entity_type="gene" role="component" text="apg4" ext_ids="855498" norm_text="" ext_dbs="NCBI gene" in_caption="True" ext_names="ATG4" ext_tax_ids="559292" ext_tax_names="Saccharomyces cerevisiae S288C" ext_urls="http://www.ncbi.nlm.nih.gov/gene/">apg4</sd-tag></em> cells co-expressing <sd-tag id="sdTag49" source="sdapi" category="entity" entity_type="gene" role="component" text="Apg7" ext_ids="856576" norm_text="" ext_dbs="NCBI gene" in_caption="True" ext_names="ATG7" ext_tax_ids="559292" ext_tax_names="Saccharomyces cerevisiae S288C" ext_urls="http://www.ncbi.nlm.nih.gov/gene/">Apg7</sd-tag>-<sd-tag id="sdTag50" source="sdapi" category="entity" entity_type="protein" role="reporter" text="Myc" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">Myc</sd-tag> and <sd-tag id="sdTag58" source="sdapi" category="entity" entity_type="protein" role="reporter" text="HA" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">HA</sd-tag>-<sd-tag id="sdTag51" source="sdapi" category="entity" entity_type="gene" role="component" text="Apg8" ext_ids="852200" norm_text="" ext_dbs="NCBI gene" in_caption="True" ext_names="ATG8" ext_tax_ids="559292" ext_tax_names="Saccharomyces cerevisiae S288C" ext_urls="http://www.ncbi.nlm.nih.gov/gene/">Apg8</sd-tag> (FGR, FG and F) from 2µ plasmids. FGR, wild-type <sd-tag id="sdTag52" source="sdapi" category="entity" entity_type="protein" role="assayed" text="Apg8" ext_ids="P38182" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="ATG8" ext_tax_ids="559292" ext_tax_names="Saccharomyces cerevisiae S288C" ext_urls="https://www.uniprot.org/uniprot/">Apg8</sd-tag>; FG, <sd-tag id="sdTag53" source="sdapi" category="entity" entity_type="protein" role="assayed" text="Apg8" ext_ids="P38182" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="ATG8" ext_tax_ids="559292" ext_tax_names="Saccharomyces cerevisiae S288C" ext_urls="https://www.uniprot.org/uniprot/">Apg8</sd-tag> lacking C-terminal arginine; F, <sd-tag id="sdTag54" source="sdapi" category="entity" entity_type="protein" role="assayed" text="Apg8" ext_ids="P38182" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="ATG8" ext_tax_ids="559292" ext_tax_names="Saccharomyces cerevisiae S288C" ext_urls="https://www.uniprot.org/uniprot/">Apg8</sd-tag> lacking C-terminal glycine and arginine. Cell lysates were <sd-tag id="sdTag55" source="sdapi" category="assay" entity_type="" role="" text="immunoprecipitated" ext_ids="BAO_0002508" norm_text="" ext_dbs="BAO" in_caption="True" ext_names="immunoprecipitation" ext_tax_ids="" ext_tax_names="" ext_urls="https://bioportal.bioontology.org/ontologies/BAO/?p=classes&conceptid=http%3A%2F%2Fwww.bioassayontology.org%2Fbao%23">immunoprecipitated</sd-tag> with anti-<sd-tag id="sdTag59" source="sdapi" category="entity" entity_type="protein" role="reporter" text="HA" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">HA</sd-tag> antibody, and <sd-tag id="sdTag56" source="sdapi" category="assay" entity_type="" role="" text="immunoblotted" ext_ids="BAO_0002424" norm_text="" ext_dbs="BAO" in_caption="True" ext_names="western blot" ext_tax_ids="" ext_tax_names="" ext_urls="https://bioportal.bioontology.org/ontologies/BAO/?p=classes&conceptid=http%3A%2F%2Fwww.bioassayontology.org%2Fbao%23">immunoblotted</sd-tag> with anti-<sd-tag id="sdTag60" source="sdapi" category="entity" entity_type="protein" role="reporter" text="HA" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">HA</sd-tag> and anti-<sd-tag id="sdTag57" source="sdapi" category="entity" entity_type="protein" role="reporter" text="Myc" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">Myc</sd-tag> antibody.</p><graphic href="https://api.sourcedata.io/file.php?panel_id=9132"/></sd-panel></fig><fig id="2634"><title></title><label>figf3</label><graphic href="https://api.sourcedata.io/file.php?figure_id=2634"/><sd-panel panel_id="9135"><strong>b</strong>, Interaction between <sd-tag id="sdTag67" source="sdapi" category="entity" entity_type="protein" role="assayed" text="Apg8" ext_ids="P38182" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="ATG8" ext_tax_ids="559292" ext_tax_names="Saccharomyces cerevisiae S288C" ext_urls="https://www.uniprot.org/uniprot/">Apg8</sd-tag> <span>and <sd-tag id="sdTag63" source="sdapi" category="entity" entity_type="protein" role="assayed" text="Apg3" ext_ids="P40344" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="ATG3" ext_tax_ids="559292" ext_tax_names="Saccharomyces cerevisiae S288C" ext_urls="https://www.uniprot.org/uniprot/">Apg3</sd-tag>. Lysates of Δ</span><em><sd-tag id="sdTag64" source="sdapi" category="entity" entity_type="gene" role="intervention" text="apg3" ext_ids="855741" norm_text="" ext_dbs="NCBI gene" in_caption="True" ext_names="ATG3" ext_tax_ids="559292" ext_tax_names="Saccharomyces cerevisiae S288C" ext_urls="http://www.ncbi.nlm.nih.gov/gene/">apg3</sd-tag></em> <span>cells expressing <sd-tag id="sdTag65" source="sdapi" category="entity" entity_type="protein" role="assayed" text="APG3" ext_ids="P40344" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="ATG3" ext_tax_ids="559292" ext_tax_names="Saccharomyces cerevisiae S288C" ext_urls="https://www.uniprot.org/uniprot/">APG3</sd-tag> and/or</span> <sd-tag id="sdTag68" source="sdapi" category="entity" entity_type="protein" role="reporter" text="Myc" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">Myc</sd-tag>-<span><sd-tag id="sdTag66" source="sdapi" category="entity" entity_type="gene" role="intervention" text="APG8" ext_ids="852200" norm_text="" ext_dbs="NCBI gene" in_caption="True" ext_names="ATG8" ext_tax_ids="559292" ext_tax_names="Saccharomyces cerevisiae S288C" ext_urls="http://www.ncbi.nlm.nih.gov/gene/">APG8</sd-tag></span> from 2µ plasmids were <sd-tag id="sdTag69" source="sdapi" category="assay" entity_type="" role="" text="immunoprecipitated" ext_ids="BAO_0002508" norm_text="" ext_dbs="BAO" in_caption="True" ext_names="immunoprecipitation" ext_tax_ids="" ext_tax_names="" ext_urls="https://bioportal.bioontology.org/ontologies/BAO/?p=classes&conceptid=http%3A%2F%2Fwww.bioassayontology.org%2Fbao%23">immunoprecipitated</sd-tag> <span>with anti-<sd-tag id="sdTag70" source="sdapi" category="entity" entity_type="protein" role="assayed" text="Apg3" ext_ids="P40344" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="ATG3" ext_tax_ids="559292" ext_tax_names="Saccharomyces cerevisiae S288C" ext_urls="https://www.uniprot.org/uniprot/">Apg3</sd-tag> antibody, and</span> <sd-tag id="sdTag71" source="sdapi" category="assay" entity_type="" role="" text="immunoblotted" ext_ids="BAO_0002424" norm_text="" ext_dbs="BAO" in_caption="True" ext_names="western blot" ext_tax_ids="" ext_tax_names="" ext_urls="https://bioportal.bioontology.org/ontologies/BAO/?p=classes&conceptid=http%3A%2F%2Fwww.bioassayontology.org%2Fbao%23">immunoblotted</sd-tag> <span>with anti-<sd-tag id="sdTag72" source="sdapi" category="entity" entity_type="protein" role="assayed" text="Apg3" ext_ids="P40344" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="ATG3" ext_tax_ids="559292" ext_tax_names="Saccharomyces cerevisiae S288C" ext_urls="https://www.uniprot.org/uniprot/">Apg3</sd-tag> and anti-</span><sd-tag id="sdTag73" source="sdapi" category="entity" entity_type="protein" role="reporter" text="Myc" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">Myc</sd-tag> antibody.<graphic href="https://api.sourcedata.io/file.php?panel_id=9135"/></sd-panel><sd-panel panel_id="9136"><strong>c</strong>, Effect of mutated Cys 234 in <span><sd-tag id="sdTag75" source="sdapi" category="entity" entity_type="gene" role="intervention" text="Apg3" ext_ids="855741" norm_text="" ext_dbs="NCBI gene" in_caption="True" ext_names="ATG3" ext_tax_ids="559292" ext_tax_names="Saccharomyces cerevisiae S288C" ext_urls="http://www.ncbi.nlm.nih.gov/gene/">Apg3</sd-tag></span> on the interaction between <sd-tag id="sdTag76" source="sdapi" category="entity" entity_type="protein" role="assayed" text="Apg8" ext_ids="P38182" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="ATG8" ext_tax_ids="559292" ext_tax_names="Saccharomyces cerevisiae S288C" ext_urls="https://www.uniprot.org/uniprot/">Apg8</sd-tag> and <sd-tag id="sdTag77" source="sdapi" category="entity" entity_type="protein" role="assayed" text="Apg3" ext_ids="P40344" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="ATG3" ext_tax_ids="559292" ext_tax_names="Saccharomyces cerevisiae S288C" ext_urls="https://www.uniprot.org/uniprot/">Apg3</sd-tag>. Lysates of Δ<em><sd-tag id="sdTag78" source="sdapi" category="entity" entity_type="gene" role="intervention" text="apg3" ext_ids="855741" norm_text="" ext_dbs="NCBI gene" in_caption="True" ext_names="ATG3" ext_tax_ids="559292" ext_tax_names="Saccharomyces cerevisiae S288C" ext_urls="http://www.ncbi.nlm.nih.gov/gene/">apg3</sd-tag></em> cells expressing <sd-tag id="sdTag79" source="sdapi" category="entity" entity_type="protein" role="reporter" text="Myc" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">Myc</sd-tag>-<sd-tag id="sdTag80" source="sdapi" category="entity" entity_type="protein" role="assayed" text="APG8" ext_ids="P38182" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="ATG8" ext_tax_ids="559292" ext_tax_names="Saccharomyces cerevisiae S288C" ext_urls="https://www.uniprot.org/uniprot/">APG8</sd-tag> and either wild-type (wt) <sd-tag id="sdTag81" source="sdapi" category="entity" entity_type="protein" role="assayed" text="APG3" ext_ids="P40344" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="ATG3" ext_tax_ids="559292" ext_tax_names="Saccharomyces cerevisiae S288C" ext_urls="https://www.uniprot.org/uniprot/">APG3</sd-tag> or mutant <sd-tag id="sdTag82" source="sdapi" category="entity" entity_type="protein" role="assayed" text="APG3" ext_ids="P40344" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="ATG3" ext_tax_ids="559292" ext_tax_names="Saccharomyces cerevisiae S288C" ext_urls="https://www.uniprot.org/uniprot/">APG3</sd-tag> (C234S and C234A) from 2µ plasmids were <sd-tag id="sdTag83" source="sdapi" category="assay" entity_type="" role="" text="immunoprecipitated" ext_ids="BAO_0002508" norm_text="" ext_dbs="BAO" in_caption="True" ext_names="immunoprecipitation" ext_tax_ids="" ext_tax_names="" ext_urls="https://bioportal.bioontology.org/ontologies/BAO/?p=classes&conceptid=http%3A%2F%2Fwww.bioassayontology.org%2Fbao%23">immunoprecipitated</sd-tag> with anti-<sd-tag id="sdTag84" source="sdapi" category="entity" entity_type="protein" role="reporter" text="Myc" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">Myc</sd-tag> antibody, and <sd-tag id="sdTag85" source="sdapi" category="assay" entity_type="" role="" text="immunoblotted" ext_ids="BAO_0002424" norm_text="" ext_dbs="BAO" in_caption="True" ext_names="western blot" ext_tax_ids="" ext_tax_names="" ext_urls="https://bioportal.bioontology.org/ontologies/BAO/?p=classes&conceptid=http%3A%2F%2Fwww.bioassayontology.org%2Fbao%23">immunoblotted</sd-tag> with anti-<sd-tag id="sdTag86" source="sdapi" category="entity" entity_type="protein" role="reporter" text="Myc" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">Myc</sd-tag> and anti-<sd-tag id="sdTag87" source="sdapi" category="entity" entity_type="protein" role="assayed" text="Apg3" ext_ids="P40344" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="ATG3" ext_tax_ids="559292" ext_tax_names="Saccharomyces cerevisiae S288C" ext_urls="https://www.uniprot.org/uniprot/">Apg3</sd-tag> antibodies.<graphic href="https://api.sourcedata.io/file.php?panel_id=9136"/></sd-panel><sd-panel panel_id="9137"><strong>d</strong><span>, <sd-tag id="sdTag89" source="sdapi" category="entity" entity_type="molecule" role="component" text="Thioester" ext_ids="CHEBI:51277" norm_text="" ext_dbs="ChEBI" in_caption="True" ext_names="thioester" ext_tax_ids="" ext_tax_names="" ext_urls="http://www.ebi.ac.uk/chebi/searchId.do?chebiId=">Thioester</sd-tag> formation between</span> <sd-tag id="sdTag93" source="sdapi" category="entity" entity_type="protein" role="assayed" text="Apg8" ext_ids="P38182" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="ATG8" ext_tax_ids="559292" ext_tax_names="Saccharomyces cerevisiae S288C" ext_urls="https://www.uniprot.org/uniprot/">Apg8</sd-tag> and <sd-tag id="sdTag94" source="sdapi" category="entity" entity_type="protein" role="assayed" text="Apg3" ext_ids="P40344" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="ATG3" ext_tax_ids="559292" ext_tax_names="Saccharomyces cerevisiae S288C" ext_urls="https://www.uniprot.org/uniprot/">Apg3</sd-tag>. Δ<em><sd-tag id="sdTag95" source="sdapi" category="entity" entity_type="gene" role="intervention" text="apg3" ext_ids="855741" norm_text="" ext_dbs="NCBI gene" in_caption="True" ext_names="ATG3" ext_tax_ids="559292" ext_tax_names="Saccharomyces cerevisiae S288C" ext_urls="http://www.ncbi.nlm.nih.gov/gene/">apg3</sd-tag></em>, Δ<em><sd-tag id="sdTag96" source="sdapi" category="entity" entity_type="gene" role="intervention" text="apg7" ext_ids="856576" norm_text="" ext_dbs="NCBI gene" in_caption="True" ext_names="ATG7" ext_tax_ids="559292" ext_tax_names="Saccharomyces cerevisiae S288C" ext_urls="http://www.ncbi.nlm.nih.gov/gene/">apg7</sd-tag></em> and Δ<em><sd-tag id="sdTag90" source="sdapi" category="entity" entity_type="gene" role="intervention" text="apg10" ext_ids="850684" norm_text="" ext_dbs="NCBI gene" in_caption="True" ext_names="ATG10" ext_tax_ids="559292" ext_tax_names="Saccharomyces cerevisiae S288C" ext_urls="http://www.ncbi.nlm.nih.gov/gene/">apg10</sd-tag></em> cells were transformed with both <sd-tag id="sdTag97" source="sdapi" category="entity" entity_type="protein" role="reporter" text="Myc" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">Myc</sd-tag>-<sd-tag id="sdTag98" source="sdapi" category="entity" entity_type="gene" role="component" text="APG8" ext_ids="852200" norm_text="" ext_dbs="NCBI gene" in_caption="True" ext_names="ATG8" ext_tax_ids="559292" ext_tax_names="Saccharomyces cerevisiae S288C" ext_urls="http://www.ncbi.nlm.nih.gov/gene/">APG8</sd-tag> and <sd-tag id="sdTag99" source="sdapi" category="entity" entity_type="gene" role="component" text="Apg3" ext_ids="855741" norm_text="" ext_dbs="NCBI gene" in_caption="True" ext_names="ATG3" ext_tax_ids="559292" ext_tax_names="Saccharomyces cerevisiae S288C" ext_urls="http://www.ncbi.nlm.nih.gov/gene/">Apg3</sd-tag>, or only <sd-tag id="sdTag100" source="sdapi" category="entity" entity_type="protein" role="assayed" text="Apg3" ext_ids="P40344" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="ATG3" ext_tax_ids="559292" ext_tax_names="Saccharomyces cerevisiae S288C" ext_urls="https://www.uniprot.org/uniprot/">Apg3</sd-tag>, on 2µ plasmids. Cell extracts were <sd-tag id="sdTag101" source="sdapi" category="assay" entity_type="" role="" text="immunoprecipitated" ext_ids="BAO_0002508" norm_text="" ext_dbs="BAO" in_caption="True" ext_names="immunoprecipitation" ext_tax_ids="" ext_tax_names="" ext_urls="https://bioportal.bioontology.org/ontologies/BAO/?p=classes&conceptid=http%3A%2F%2Fwww.bioassayontology.org%2Fbao%23">immunoprecipitated</sd-tag> with anti-<sd-tag id="sdTag102" source="sdapi" category="entity" entity_type="protein" role="reporter" text="Myc" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">Myc</sd-tag> <span>antibody, and subjected to <sd-tag id="sdTag91" source="sdapi" category="assay" entity_type="" role="" text="immunoblot" ext_ids="BAO_0002424" norm_text="" ext_dbs="BAO" in_caption="True" ext_names="western blot" ext_tax_ids="" ext_tax_names="" ext_urls="https://bioportal.bioontology.org/ontologies/BAO/?p=classes&conceptid=http%3A%2F%2Fwww.bioassayontology.org%2Fbao%23">immunoblot</sd-tag> (IB) using anti-</span><sd-tag id="sdTag103" source="sdapi" category="entity" entity_type="protein" role="reporter" text="Myc" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">Myc</sd-tag> and anti-<sd-tag id="sdTag104" source="sdapi" category="entity" entity_type="protein" role="assayed" text="Apg3" ext_ids="P40344" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="ATG3" ext_tax_ids="559292" ext_tax_names="Saccharomyces cerevisiae S288C" ext_urls="https://www.uniprot.org/uniprot/">Apg3</sd-tag> <span>antibody. <sd-tag id="sdTag92" source="sdapi" category="entity" entity_type="molecule" role="intervention" text="DTT" ext_ids="CHEBI:18320" norm_text="" ext_dbs="ChEBI" in_caption="True" ext_names="1,4-dithiothreitol" ext_tax_ids="" ext_tax_names="" ext_urls="http://www.ebi.ac.uk/chebi/searchId.do?chebiId=">DTT</sd-tag>-, no <sd-tag id="sdTag105" source="sdapi" category="entity" entity_type="molecule" role="intervention" text="DTT" ext_ids="CHEBI:18320" norm_text="" ext_dbs="ChEBI" in_caption="True" ext_names="1,4-dithiothreitol" ext_tax_ids="" ext_tax_names="" ext_urls="http://www.ebi.ac.uk/chebi/searchId.do?chebiId=">DTT</sd-tag>; <sd-tag id="sdTag106" source="sdapi" category="entity" entity_type="molecule" role="intervention" text="DTT" ext_ids="CHEBI:18320" norm_text="" ext_dbs="ChEBI" in_caption="True" ext_names="1,4-dithiothreitol" ext_tax_ids="" ext_tax_names="" ext_urls="http://www.ebi.ac.uk/chebi/searchId.do?chebiId=">DTT</sd-tag>+, 100 mM <sd-tag id="sdTag107" source="sdapi" category="entity" entity_type="molecule" role="intervention" text="DTT" ext_ids="CHEBI:18320" norm_text="" ext_dbs="ChEBI" in_caption="True" ext_names="1,4-dithiothreitol" ext_tax_ids="" ext_tax_names="" ext_urls="http://www.ebi.ac.uk/chebi/searchId.do?chebiId=">DTT</sd-tag>; asterisk,</span> <sd-tag id="sdTag108" source="sdapi" category="entity" entity_type="protein" role="assayed" text="Apg8" ext_ids="P38182" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="ATG8" ext_tax_ids="559292" ext_tax_names="Saccharomyces cerevisiae S288C" ext_urls="https://www.uniprot.org/uniprot/">Apg8</sd-tag>-<sd-tag id="sdTag109" source="sdapi" category="entity" entity_type="protein" role="assayed" text="Apg3" ext_ids="P40344" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="ATG3" ext_tax_ids="559292" ext_tax_names="Saccharomyces cerevisiae S288C" ext_urls="https://www.uniprot.org/uniprot/">Apg3</sd-tag> conjugate; diamond, possible <sd-tag id="sdTag110" source="sdapi" category="entity" entity_type="protein" role="reporter" text="Myc" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">Myc</sd-tag>-<sd-tag id="sdTag111" source="sdapi" category="entity" entity_type="protein" role="assayed" text="Apg8" ext_ids="P38182" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="ATG8" ext_tax_ids="559292" ext_tax_names="Saccharomyces cerevisiae S288C" ext_urls="https://www.uniprot.org/uniprot/">Apg8</sd-tag> dimer.<graphic href="https://api.sourcedata.io/file.php?panel_id=9137"/></sd-panel></fig><fig id="2635"><title></title><label>figf4</label><graphic href="https://api.sourcedata.io/file.php?figure_id=2635"/><sd-panel panel_id="9138"><strong>a</strong>, <span><sd-tag id="sdTag120" source="sdapi" category="entity" entity_type="gene" role="intervention" text="Apg7" ext_ids="856576" norm_text="" ext_dbs="NCBI gene" in_caption="True" ext_names="ATG7" ext_tax_ids="559292" ext_tax_names="Saccharomyces cerevisiae S288C" ext_urls="http://www.ncbi.nlm.nih.gov/gene/">Apg7</sd-tag></span> and <span><sd-tag id="sdTag121" source="sdapi" category="entity" entity_type="gene" role="intervention" text="Apg3" ext_ids="855741" norm_text="" ext_dbs="NCBI gene" in_caption="True" ext_names="ATG3" ext_tax_ids="559292" ext_tax_names="Saccharomyces cerevisiae S288C" ext_urls="http://www.ncbi.nlm.nih.gov/gene/">Apg3</sd-tag></span> are necessary for attachment of <sd-tag id="sdTag117" source="sdapi" category="entity" entity_type="protein" role="assayed" text="Apg8" ext_ids="P38182///A6ZKM4" norm_text="" ext_dbs="Uniprot///Uniprot" in_caption="True" ext_names="ATG8///ATG8" ext_tax_ids="307796///559292" ext_tax_names="Saccharomyces cerevisiae YJM789///Saccharomyces cerevisiae S288C" ext_urls="https://www.uniprot.org/uniprot////https://www.uniprot.org/uniprot/">Apg8</sd-tag> <span>to <sd-tag id="sdTag158" source="sdapi" category="entity" entity_type="molecule" role="component" text="PE" ext_ids="CHEBI:16038" norm_text="" ext_dbs="ChEBI" in_caption="True" ext_names="phosphatidylethanolamine" ext_tax_ids="" ext_tax_names="" ext_urls="http://www.ebi.ac.uk/chebi/searchId.do?chebiId=">PE</sd-tag>. Δ</span><em>apg</em> <span>cell lysates were subjected to <sd-tag id="sdTag113" source="sdapi" category="assay" entity_type="" role="" text="SDS–PAGE" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">SDS-PAGE</sd-tag> containing 6 M <sd-tag id="sdTag114" source="sdapi" category="entity" entity_type="molecule" role="component" text="urea" ext_ids="CHEBI:16199" norm_text="" ext_dbs="ChEBI" in_caption="True" ext_names="urea" ext_tax_ids="" ext_tax_names="" ext_urls="http://www.ebi.ac.uk/chebi/searchId.do?chebiId=">urea</sd-tag>, and</span> <sd-tag id="sdTag118" source="sdapi" category="assay" entity_type="" role="" text="immunoblotted" ext_ids="BAO_0002424" norm_text="" ext_dbs="BAO" in_caption="True" ext_names="western blot" ext_tax_ids="" ext_tax_names="" ext_urls="https://bioportal.bioontology.org/ontologies/BAO/?p=classes&conceptid=http%3A%2F%2Fwww.bioassayontology.org%2Fbao%23">immunoblotted</sd-tag> with anti-<sd-tag id="sdTag119" source="sdapi" category="entity" entity_type="protein" role="assayed" text="Apg8" ext_ids="P38182///A6ZKM4" norm_text="" ext_dbs="Uniprot///Uniprot" in_caption="True" ext_names="ATG8///ATG8" ext_tax_ids="307796///559292" ext_tax_names="Saccharomyces cerevisiae YJM789///Saccharomyces cerevisiae S288C" ext_urls="https://www.uniprot.org/uniprot////https://www.uniprot.org/uniprot/">Apg8</sd-tag> antibody.<graphic href="https://api.sourcedata.io/file.php?panel_id=9138"/></sd-panel><sd-panel panel_id="9139"><strong>b</strong>, Autophagic activity in <em><sd-tag id="sdTag127" source="sdapi" category="entity" entity_type="gene" role="intervention" text="apg3" ext_ids="855741" norm_text="" ext_dbs="NCBI gene" in_caption="True" ext_names="ATG3" ext_tax_ids="559292" ext_tax_names="Saccharomyces cerevisiae S288C" ext_urls="http://www.ncbi.nlm.nih.gov/gene/">apg3</sd-tag></em> mutants. Wild-type, Δ<em><sd-tag id="sdTag128" source="sdapi" category="entity" entity_type="gene" role="intervention" text="apg3" ext_ids="855741" norm_text="" ext_dbs="NCBI gene" in_caption="True" ext_names="ATG3" ext_tax_ids="559292" ext_tax_names="Saccharomyces cerevisiae S288C" ext_urls="http://www.ncbi.nlm.nih.gov/gene/">apg3</sd-tag></em> and <em><sd-tag id="sdTag129" source="sdapi" category="entity" entity_type="gene" role="intervention" text="apg3" ext_ids="855741" norm_text="" ext_dbs="NCBI gene" in_caption="True" ext_names="ATG3" ext_tax_ids="559292" ext_tax_names="Saccharomyces cerevisiae S288C" ext_urls="http://www.ncbi.nlm.nih.gov/gene/">apg3</sd-tag></em><span>C243A cells were grown in rich medium (open bar) and subjected to <sd-tag id="sdTag124" source="sdapi" category="entity" entity_type="molecule" role="intervention" text="nitrogen" ext_ids="CHEBI:25555" norm_text="" ext_dbs="ChEBI" in_caption="True" ext_names="nitrogen atom" ext_tax_ids="" ext_tax_names="" ext_urls="http://www.ebi.ac.uk/chebi/searchId.do?chebiId=">nitrogen</sd-tag> starvation for <sd-tag id="sdTag125" source="sdapi" category="time" entity_type="" role="" text="4 h" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">4 h</sd-tag> (filled bar). The autophagic activity of each cell was assessed by measuring the <sd-tag id="sdTag126" source="sdapi" category="entity" entity_type="subcellular" role="assayed" text="alkaline phosphatase" ext_ids="GO:1903293" norm_text="" ext_dbs="Gene Ontology" in_caption="True" ext_names="phosphatase complex" ext_tax_ids="" ext_tax_names="" ext_urls="http://amigo.geneontology.org/amigo/term/">alkaline phosphatase</sd-tag> activity. The</span> <em><sd-tag id="sdTag130" source="sdapi" category="entity" entity_type="gene" role="intervention" text="apg3" ext_ids="855741" norm_text="" ext_dbs="NCBI gene" in_caption="True" ext_names="ATG3" ext_tax_ids="559292" ext_tax_names="Saccharomyces cerevisiae S288C" ext_urls="http://www.ncbi.nlm.nih.gov/gene/">apg3</sd-tag></em>C243A mutant showed defective autophagy.<graphic href="https://api.sourcedata.io/file.php?panel_id=9139"/></sd-panel></fig></article> |
