token_classification/10-1016_j-celrep-2015-06-022.xml

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<article doi="10.1016/j.celrep.2015.06.022" abstract="Communication between organelles is crucial for eukaryotic cells to function as one coherent unit. An important means of communication is through membrane contact sites, where two organelles come into close proximity allowing the transport of lipids and small solutes between them. Contact sites are dynamic in size and can change in response to environmental or cellular stimuli; however, how this is regulated has been unclear. Here, we show that Saccharomyces cerevisiae Lam6 resides in several central contact sites: ERMES (ER/mitochondria encounter structure), vCLAMP (vacuole and mitochondria patch), and NVJ (nuclear vacuolar junction). We show that Lam6 is sufficient for expansion of contact sites under physiological conditions and necessary for coordination of contact site size. Given that Lam6 is part of a large protein family and is conserved in vertebrates, our work opens avenues for investigating the underlying principles of organelle communication."><fig id="5019"><title></title><label>Figure 1</label><graphic href="https://api.sourcedata.io/file.php?figure_id=5019"/><sd-panel panel_id="15299"><p>(G). <sd-tag id="sdTag1" source="sdapi" category="entity" entity_type="protein" role="reporter" text="GFP" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">GFP</sd-tag>-tagged <sd-tag id="sdTag2" source="sdapi" category="entity" entity_type="subcellular" role="component" text="ERMES" ext_ids="GO:0032865" norm_text="" ext_dbs="Gene Ontology" in_caption="True" ext_names="ERMES complex" ext_tax_ids="" ext_tax_names="" ext_urls="http://amigo.geneontology.org/amigo/term/">ERMES</sd-tag> complex proteins retain their characteristic punctate structure in Δ<sd-tag id="sdTag3" source="sdapi" category="entity" entity_type="gene" role="component" text="lam6" ext_ids="850761" norm_text="" ext_dbs="NCBI gene" in_caption="True" ext_names="LAM6" ext_tax_ids="559292" ext_tax_names="Saccharomyces cerevisiae S288C" ext_urls="http://www.ncbi.nlm.nih.gov/gene/">lam6</sd-tag>, suggesting that <sd-tag id="sdTag4" source="sdapi" category="entity" entity_type="gene" role="component" text="Lam6" ext_ids="850761" norm_text="" ext_dbs="NCBI gene" in_caption="True" ext_names="LAM6" ext_tax_ids="559292" ext_tax_names="Saccharomyces cerevisiae S288C" ext_urls="http://www.ncbi.nlm.nih.gov/gene/">Lam6</sd-tag> is not an essential complex member. Scale bar represents 5 μm.</p><graphic href="https://api.sourcedata.io/file.php?panel_id=15299"/></sd-panel></fig><fig id="5020"><title></title><label>Figure 2</label><graphic href="https://api.sourcedata.io/file.php?figure_id=5020"/><sd-panel panel_id="15300"><span>(A) <sd-tag id="sdTag11" source="sdapi" category="assay" entity_type="" role="" text="Fluorescence microscopy" ext_ids="BAO_0000450" norm_text="" ext_dbs="BAO" in_caption="True" ext_names="fluorescence microscopy" ext_tax_ids="" ext_tax_names="" ext_urls="https://bioportal.bioontology.org/ontologies/BAO/?p=classes&amp;conceptid=http%3A%2F%2Fwww.bioassayontology.org%2Fbao%23">Fluorescence microscopy</sd-tag> demonstrates that</span> <span><sd-tag id="sdTag12" source="sdapi" category="entity" entity_type="protein" role="assayed" text="Lam6" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">Lam6</sd-tag></span>-<sd-tag id="sdTag20" source="sdapi" category="entity" entity_type="protein" role="reporter" text="GFP" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">GFP</sd-tag> co-localizes with <sd-tag id="sdTag21" source="sdapi" category="entity" entity_type="subcellular" role="component" text="ERMES" ext_ids="GO:0032865" norm_text="" ext_dbs="Gene Ontology" in_caption="True" ext_names="ERMES complex" ext_tax_ids="" ext_tax_names="" ext_urls="http://amigo.geneontology.org/amigo/term/">ERMES</sd-tag> <span>(<sd-tag id="sdTag13" source="sdapi" category="entity" entity_type="protein" role="assayed" text="Mdm34" ext_ids="P53083" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="MDM34" ext_tax_ids="559292" ext_tax_names="Saccharomyces cerevisiae S288C" ext_urls="https://www.uniprot.org/uniprot/">Mdm34</sd-tag>-<sd-tag id="sdTag14" source="sdapi" category="entity" entity_type="protein" role="reporter" text="Cherry" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">Cherry</sd-tag>) (yellow arrows) and also localizes to non-</span><sd-tag id="sdTag22" source="sdapi" category="entity" entity_type="subcellular" role="component" text="ERMES" ext_ids="GO:0032865" norm_text="" ext_dbs="Gene Ontology" in_caption="True" ext_names="ERMES complex" ext_tax_ids="" ext_tax_names="" ext_urls="http://amigo.geneontology.org/amigo/term/">ERMES</sd-tag> <span>locations in the cell (red arrows). These additional locations co-localized with the <sd-tag id="sdTag15" source="sdapi" category="entity" entity_type="subcellular" role="component" text="NVJ" ext_ids="GO:0071561" norm_text="" ext_dbs="Gene Ontology" in_caption="True" ext_names="nucleus-vacuole junction" ext_tax_ids="" ext_tax_names="" ext_urls="http://amigo.geneontology.org/amigo/term/">NVJ</sd-tag> (<sd-tag id="sdTag16" source="sdapi" category="entity" entity_type="protein" role="assayed" text="Nvj1" ext_ids="P38881" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="NVJ1" ext_tax_ids="559292" ext_tax_names="Saccharomyces cerevisiae S288C" ext_urls="https://www.uniprot.org/uniprot/">Nvj1</sd-tag>-</span><sd-tag id="sdTag23" source="sdapi" category="entity" entity_type="protein" role="reporter" text="GFP" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">GFP</sd-tag><span>) as well as with the <sd-tag id="sdTag17" source="sdapi" category="entity" entity_type="subcellular" role="component" text="vCLAMP" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">vCLAMP</sd-tag> (</span><sd-tag id="sdTag24" source="sdapi" category="entity" entity_type="protein" role="reporter" text="GFP" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">GFP</sd-tag><span>-<sd-tag id="sdTag18" source="sdapi" category="entity" entity_type="protein" role="assayed" text="Vps39" ext_ids="Q07468" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="VAM6" ext_tax_ids="559292" ext_tax_names="Saccharomyces cerevisiae S288C" ext_urls="https://www.uniprot.org/uniprot/">Vps39</sd-tag>). Scale bar represents 5 μm.</span><graphic href="https://api.sourcedata.io/file.php?panel_id=15300"/></sd-panel><sd-panel panel_id="15301">(B) Overexpression and tagging of <sd-tag id="sdTag25" source="sdapi" category="entity" entity_type="protein" role="assayed" text="Lam6" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">Lam6</sd-tag> confirmed that it co-localizes with <sd-tag id="sdTag26" source="sdapi" category="entity" entity_type="subcellular" role="component" text="ERMES" ext_ids="GO:0032865" norm_text="" ext_dbs="Gene Ontology" in_caption="True" ext_names="ERMES complex" ext_tax_ids="" ext_tax_names="" ext_urls="http://amigo.geneontology.org/amigo/term/">ERMES</sd-tag> (<sd-tag id="sdTag27" source="sdapi" category="entity" entity_type="protein" role="assayed" text="Mdm34" ext_ids="P53083" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="MDM34" ext_tax_ids="559292" ext_tax_names="Saccharomyces cerevisiae S288C" ext_urls="https://www.uniprot.org/uniprot/">Mdm34</sd-tag>-<sd-tag id="sdTag28" source="sdapi" category="entity" entity_type="protein" role="reporter" text="GFP" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">GFP</sd-tag>) (yellow arrows) as well as to additional contact sites (red arrows), <sd-tag id="sdTag29" source="sdapi" category="entity" entity_type="subcellular" role="component" text="NVJ" ext_ids="GO:0071561" norm_text="" ext_dbs="Gene Ontology" in_caption="True" ext_names="nucleus-vacuole junction" ext_tax_ids="" ext_tax_names="" ext_urls="http://amigo.geneontology.org/amigo/term/">NVJ</sd-tag> (marked by <sd-tag id="sdTag30" source="sdapi" category="entity" entity_type="protein" role="assayed" text="Nvj1" ext_ids="P38881" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="NVJ1" ext_tax_ids="559292" ext_tax_names="Saccharomyces cerevisiae S288C" ext_urls="https://www.uniprot.org/uniprot/">Nvj1</sd-tag>) and <sd-tag id="sdTag31" source="sdapi" category="entity" entity_type="subcellular" role="component" text="vCLAMP" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">vCLAMP</sd-tag> (marked by <sd-tag id="sdTag32" source="sdapi" category="entity" entity_type="protein" role="assayed" text="Vps39" ext_ids="Q07468" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="VAM6" ext_tax_ids="559292" ext_tax_names="Saccharomyces cerevisiae S288C" ext_urls="https://www.uniprot.org/uniprot/">Vps39</sd-tag>). Scale bar represents 5 μm.<graphic href="https://api.sourcedata.io/file.php?panel_id=15301"/></sd-panel></fig><fig id="5021"><title></title><label>Figure 3</label><graphic href="https://api.sourcedata.io/file.php?figure_id=5021"/><sd-panel panel_id="15302">(A) <sd-tag id="sdTag37" source="sdapi" category="assay" entity_type="" role="" text="Fluorescence microscopy" ext_ids="BAO_0000450" norm_text="" ext_dbs="BAO" in_caption="True" ext_names="fluorescence microscopy" ext_tax_ids="" ext_tax_names="" ext_urls="https://bioportal.bioontology.org/ontologies/BAO/?p=classes&amp;conceptid=http%3A%2F%2Fwww.bioassayontology.org%2Fbao%23">Fluorescence microscopy</sd-tag> demonstrates that the overexpression of <sd-tag id="sdTag38" source="sdapi" category="entity" entity_type="protein" role="reporter" text="Cherry" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">Cherry</sd-tag>-<span><sd-tag id="sdTag36" source="sdapi" category="entity" entity_type="protein" role="assayed" text="Lam6" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">Lam6</sd-tag></span> (OE-<sd-tag id="sdTag39" source="sdapi" category="entity" entity_type="protein" role="reporter" text="Cherry" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">Cherry</sd-tag><span>-<sd-tag id="sdTag50" source="sdapi" category="entity" entity_type="protein" role="assayed" text="LAM6" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">LAM6</sd-tag>) results in an expansion of the following three contact sites:</span> <sd-tag id="sdTag40" source="sdapi" category="entity" entity_type="subcellular" role="component" text="ERMES" ext_ids="GO:0032865" norm_text="" ext_dbs="Gene Ontology" in_caption="True" ext_names="ERMES complex" ext_tax_ids="" ext_tax_names="" ext_urls="http://amigo.geneontology.org/amigo/term/">ERMES</sd-tag> (<sd-tag id="sdTag41" source="sdapi" category="entity" entity_type="protein" role="assayed" text="Mdm34" ext_ids="P53083" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="MDM34" ext_tax_ids="559292" ext_tax_names="Saccharomyces cerevisiae S288C" ext_urls="https://www.uniprot.org/uniprot/">Mdm34</sd-tag>-<sd-tag id="sdTag42" source="sdapi" category="entity" entity_type="protein" role="reporter" text="GFP" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">GFP</sd-tag>), <sd-tag id="sdTag43" source="sdapi" category="entity" entity_type="subcellular" role="component" text="NVJ" ext_ids="GO:0071561" norm_text="" ext_dbs="Gene Ontology" in_caption="True" ext_names="nucleus-vacuole junction" ext_tax_ids="" ext_tax_names="" ext_urls="http://amigo.geneontology.org/amigo/term/">NVJ</sd-tag> (<sd-tag id="sdTag44" source="sdapi" category="entity" entity_type="protein" role="assayed" text="Nvj1" ext_ids="P38881" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="NVJ1" ext_tax_ids="559292" ext_tax_names="Saccharomyces cerevisiae S288C" ext_urls="https://www.uniprot.org/uniprot/">Nvj1</sd-tag>-<sd-tag id="sdTag45" source="sdapi" category="entity" entity_type="protein" role="reporter" text="GFP" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">GFP</sd-tag>), and <sd-tag id="sdTag46" source="sdapi" category="entity" entity_type="subcellular" role="component" text="vCLAMP" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">vCLAMP</sd-tag> (<sd-tag id="sdTag47" source="sdapi" category="entity" entity_type="protein" role="reporter" text="GFP" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">GFP</sd-tag>-<sd-tag id="sdTag48" source="sdapi" category="entity" entity_type="protein" role="assayed" text="Vps39" ext_ids="Q07468" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="VAM6" ext_tax_ids="559292" ext_tax_names="Saccharomyces cerevisiae S288C" ext_urls="https://www.uniprot.org/uniprot/">Vps39</sd-tag>). This suggests that an increase in <sd-tag id="sdTag49" source="sdapi" category="entity" entity_type="gene" role="intervention" text="Lam6" ext_ids="850761" norm_text="" ext_dbs="NCBI gene" in_caption="True" ext_names="LAM6" ext_tax_ids="559292" ext_tax_names="Saccharomyces cerevisiae S288C" ext_urls="http://www.ncbi.nlm.nih.gov/gene/">Lam6</sd-tag> levels in the contact site is sufficient for its expansion. The numbers represent the average contact site size (120 cells per sample). Scale bar represents 5 μm.<graphic href="https://api.sourcedata.io/file.php?panel_id=15302"/></sd-panel><sd-panel panel_id="15303"><span>(B) <sd-tag id="sdTag52" source="sdapi" category="assay" entity_type="" role="" text="Immuno-EM" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">Immuno-EM</sd-tag> verified that</span> <sd-tag id="sdTag57" source="sdapi" category="entity" entity_type="protein" role="reporter" text="GFP" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">GFP</sd-tag>-<sd-tag id="sdTag58" source="sdapi" category="entity" entity_type="gene" role="intervention" text="Lam6" ext_ids="850761" norm_text="" ext_dbs="NCBI gene" in_caption="True" ext_names="LAM6" ext_tax_ids="559292" ext_tax_names="Saccharomyces cerevisiae S288C" ext_urls="http://www.ncbi.nlm.nih.gov/gene/">Lam6</sd-tag> overexpression indeed causes an expansion of contact site size. While <sd-tag id="sdTag59" source="sdapi" category="entity" entity_type="subcellular" role="assayed" text="vCLAMP" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">vCLAMP</sd-tag> <span>was hardly visible in WT cells, it could be detected easily in the OE strain (albeit often had <sd-tag id="sdTag53" source="sdapi" category="entity" entity_type="subcellular" role="assayed" text="ER" ext_ids="GO:0005783" norm_text="" ext_dbs="Gene Ontology" in_caption="True" ext_names="endoplasmic reticulum" ext_tax_ids="" ext_tax_names="" ext_urls="http://amigo.geneontology.org/amigo/term/">ER</sd-tag> tubules invading it). The</span> <sd-tag id="sdTag60" source="sdapi" category="entity" entity_type="subcellular" role="assayed" text="NVJ" ext_ids="GO:0071561" norm_text="" ext_dbs="Gene Ontology" in_caption="True" ext_names="nucleus-vacuole junction" ext_tax_ids="" ext_tax_names="" ext_urls="http://amigo.geneontology.org/amigo/term/">NVJ</sd-tag> <span>underwent expansion as well as evoked PMN in OE strains, and the <sd-tag id="sdTag61" source="sdapi" category="entity" entity_type="subcellular" role="assayed" text="ER" ext_ids="GO:0005783" norm_text="" ext_dbs="Gene Ontology" in_caption="True" ext_names="endoplasmic reticulum" ext_tax_ids="" ext_tax_names="" ext_urls="http://amigo.geneontology.org/amigo/term/">ER</sd-tag>-<sd-tag id="sdTag62" source="sdapi" category="entity" entity_type="subcellular" role="assayed" text="mitochondria" ext_ids="GO:0005739" norm_text="" ext_dbs="Gene Ontology" in_caption="True" ext_names="mitochondrion" ext_tax_ids="" ext_tax_names="" ext_urls="http://amigo.geneontology.org/amigo/term/">mitochondria</sd-tag> contact became large and elongated instead of small and distinct. N, <sd-tag id="sdTag54" source="sdapi" category="entity" entity_type="subcellular" role="assayed" text="nucleus" ext_ids="GO:0005634" norm_text="" ext_dbs="Gene Ontology" in_caption="True" ext_names="nucleus" ext_tax_ids="" ext_tax_names="" ext_urls="http://amigo.geneontology.org/amigo/term/">nucleus</sd-tag>; M, <sd-tag id="sdTag55" source="sdapi" category="entity" entity_type="subcellular" role="assayed" text="mitochondria" ext_ids="GO:0005739" norm_text="" ext_dbs="Gene Ontology" in_caption="True" ext_names="mitochondrion" ext_tax_ids="" ext_tax_names="" ext_urls="http://amigo.geneontology.org/amigo/term/">mitochondria</sd-tag>; V, <sd-tag id="sdTag56" source="sdapi" category="entity" entity_type="subcellular" role="assayed" text="vacuole" ext_ids="GO:0005773" norm_text="" ext_dbs="Gene Ontology" in_caption="True" ext_names="vacuole" ext_tax_ids="" ext_tax_names="" ext_urls="http://amigo.geneontology.org/amigo/term/">vacuole</sd-tag>. Scale bar represents 200 nm (see also Figure S3I).</span><graphic href="https://api.sourcedata.io/file.php?panel_id=15303"/></sd-panel></fig><fig id="5022"><title></title><label>Figure 4</label><graphic href="https://api.sourcedata.io/file.php?figure_id=5022"/><sd-panel panel_id="15304"><p>(A) Fluorescent microscopy shows that the <sd-tag id="sdTag66" source="sdapi" category="entity" entity_type="subcellular" role="assayed" text="ERMES" ext_ids="GO:0032865" norm_text="" ext_dbs="Gene Ontology" in_caption="True" ext_names="ERMES complex" ext_tax_ids="" ext_tax_names="" ext_urls="http://amigo.geneontology.org/amigo/term/">ERMES</sd-tag> contact (as measured by the number of <sd-tag id="sdTag67" source="sdapi" category="entity" entity_type="protein" role="assayed" text="Mdm34" ext_ids="P53083" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="MDM34" ext_tax_ids="559292" ext_tax_names="Saccharomyces cerevisiae S288C" ext_urls="https://www.uniprot.org/uniprot/">Mdm34</sd-tag>-<sd-tag id="sdTag68" source="sdapi" category="entity" entity_type="protein" role="reporter" text="GFP" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">GFP</sd-tag> puncta per cell) expanded in the Δ<sd-tag id="sdTag65" source="sdapi" category="entity" entity_type="gene" role="intervention" text="vps39" ext_ids="851482" norm_text="" ext_dbs="NCBI gene" in_caption="True" ext_names="VAM6" ext_tax_ids="559292" ext_tax_names="Saccharomyces cerevisiae S288C" ext_urls="http://www.ncbi.nlm.nih.gov/gene/">vps39</sd-tag> background relative to WT. However, the expansion did not occur on the background of Δ<sd-tag id="sdTag69" source="sdapi" category="entity" entity_type="gene" role="intervention" text="vps39" ext_ids="851482" norm_text="" ext_dbs="NCBI gene" in_caption="True" ext_names="VAM6" ext_tax_ids="559292" ext_tax_names="Saccharomyces cerevisiae S288C" ext_urls="http://www.ncbi.nlm.nih.gov/gene/">vps39</sd-tag> Δ<sd-tag id="sdTag70" source="sdapi" category="entity" entity_type="gene" role="intervention" text="lam6" ext_ids="850761" norm_text="" ext_dbs="NCBI gene" in_caption="True" ext_names="LAM6" ext_tax_ids="559292" ext_tax_names="Saccharomyces cerevisiae S288C" ext_urls="http://www.ncbi.nlm.nih.gov/gene/">lam6</sd-tag>, demonstrating that <sd-tag id="sdTag71" source="sdapi" category="entity" entity_type="gene" role="intervention" text="Lam6" ext_ids="850761" norm_text="" ext_dbs="NCBI gene" in_caption="True" ext_names="LAM6" ext_tax_ids="559292" ext_tax_names="Saccharomyces cerevisiae S288C" ext_urls="http://www.ncbi.nlm.nih.gov/gene/">Lam6</sd-tag> is necessary for <sd-tag id="sdTag72" source="sdapi" category="entity" entity_type="subcellular" role="assayed" text="ERMES" ext_ids="GO:0032865" norm_text="" ext_dbs="Gene Ontology" in_caption="True" ext_names="ERMES complex" ext_tax_ids="" ext_tax_names="" ext_urls="http://amigo.geneontology.org/amigo/term/">ERMES</sd-tag> expansion under these conditions. Scale bar represents 5 μm.</p><p>(B) Quantitation of (A). Bars represent the percentage of cells containing the specific number of puncta/cell out of total cells counted for the strain (for WT, n = 234 cells; for Δ<sd-tag id="sdTag73" source="sdapi" category="entity" entity_type="gene" role="intervention" text="vps39" ext_ids="851482" norm_text="" ext_dbs="NCBI gene" in_caption="True" ext_names="VAM6" ext_tax_ids="559292" ext_tax_names="Saccharomyces cerevisiae S288C" ext_urls="http://www.ncbi.nlm.nih.gov/gene/">vps39</sd-tag>, n = 246; for Δ<sd-tag id="sdTag74" source="sdapi" category="entity" entity_type="gene" role="intervention" text="vps39" ext_ids="851482" norm_text="" ext_dbs="NCBI gene" in_caption="True" ext_names="VAM6" ext_tax_ids="559292" ext_tax_names="Saccharomyces cerevisiae S288C" ext_urls="http://www.ncbi.nlm.nih.gov/gene/">vps39</sd-tag> Δ<sd-tag id="sdTag75" source="sdapi" category="entity" entity_type="gene" role="intervention" text="lam6" ext_ids="850761" norm_text="" ext_dbs="NCBI gene" in_caption="True" ext_names="LAM6" ext_tax_ids="559292" ext_tax_names="Saccharomyces cerevisiae S288C" ext_urls="http://www.ncbi.nlm.nih.gov/gene/">lam6</sd-tag>, n = 271).</p><graphic href="https://api.sourcedata.io/file.php?panel_id=15304"/></sd-panel><sd-panel panel_id="15305">(C) Fluorescent microscopy demonstrates that downregulating <sd-tag id="sdTag81" source="sdapi" category="entity" entity_type="subcellular" role="component" text="ERMES" ext_ids="GO:0032865" norm_text="" ext_dbs="Gene Ontology" in_caption="True" ext_names="ERMES complex" ext_tax_ids="" ext_tax_names="" ext_urls="http://amigo.geneontology.org/amigo/term/">ERMES</sd-tag> <span>contacts (by growing <sd-tag id="sdTag78" source="sdapi" category="entity" entity_type="gene" role="component" text="GALp" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">GALp</sd-tag>-<sd-tag id="sdTag79" source="sdapi" category="entity" entity_type="gene" role="component" text="MDM34" ext_ids="852654" norm_text="" ext_dbs="NCBI gene" in_caption="True" ext_names="MDM34" ext_tax_ids="559292" ext_tax_names="Saccharomyces cerevisiae S288C" ext_urls="http://www.ncbi.nlm.nih.gov/gene/">MDM34</sd-tag> strains in <sd-tag id="sdTag80" source="sdapi" category="entity" entity_type="molecule" role="intervention" text="glucose" ext_ids="CHEBI:17234" norm_text="" ext_dbs="ChEBI" in_caption="True" ext_names="glucose" ext_tax_ids="" ext_tax_names="" ext_urls="http://www.ebi.ac.uk/chebi/searchId.do?chebiId=">glucose</sd-tag>) indeed caused expansion of</span> <sd-tag id="sdTag82" source="sdapi" category="entity" entity_type="subcellular" role="component" text="vCLAMP" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">vCLAMP</sd-tag> (<sd-tag id="sdTag83" source="sdapi" category="entity" entity_type="protein" role="reporter" text="GFP" ext_ids="" norm_text="" ext_dbs="" in_caption="True" ext_names="" ext_tax_ids="" ext_tax_names="" ext_urls="">GFP</sd-tag>-<sd-tag id="sdTag84" source="sdapi" category="entity" entity_type="protein" role="assayed" text="Vps39" ext_ids="Q07468" norm_text="" ext_dbs="Uniprot" in_caption="True" ext_names="VAM6" ext_tax_ids="559292" ext_tax_names="Saccharomyces cerevisiae S288C" ext_urls="https://www.uniprot.org/uniprot/">Vps39</sd-tag>). However, this expansion was diminished in a Δ<sd-tag id="sdTag85" source="sdapi" category="entity" entity_type="gene" role="intervention" text="lam6" ext_ids="850761" norm_text="" ext_dbs="NCBI gene" in_caption="True" ext_names="LAM6" ext_tax_ids="559292" ext_tax_names="Saccharomyces cerevisiae S288C" ext_urls="http://www.ncbi.nlm.nih.gov/gene/">lam6</sd-tag> background. Scale bar represents 5 μm (see also Figure S4).<graphic href="https://api.sourcedata.io/file.php?panel_id=15305"/></sd-panel></fig></article>