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Identification of Suppressor of Clathrin Deficiency-1 (SCD1) and Its Connection to Clathrin-Mediated Endocytosis in Saccharomyces cerevisiae
Abstract Clathrin is a major coat protein involved in vesicle formation during endocytosis and transport in the endosomal/trans Golgi system. Clathrin is required for normal growth of yeast (Saccharomyces cerevisiae) and in some genetic backgrounds deletion of the clathrin heavy chain gene (CHC1) is...
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Published in: | G3 : genes - genomes - genetics 2019-03, Vol.9 (3), p.867-877 |
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creator | Moorthy, Balaji T Sharma, Anupam Boettner, Douglas R Wilson, Thomas E Lemmon, Sandra K |
description | Abstract
Clathrin is a major coat protein involved in vesicle formation during endocytosis and transport in the endosomal/trans Golgi system. Clathrin is required for normal growth of yeast (Saccharomyces cerevisiae) and in some genetic backgrounds deletion of the clathrin heavy chain gene (CHC1) is lethal. Our lab defined a locus referred to as “suppressor of clathrin deficiency” (SCD1). In the presence of the scd1-v allele (“v” – viable), yeast cells lacking clathrin heavy chain survive but grow slowly, are morphologically abnormal and have many membrane trafficking defects. In the presence of scd1-i (“i”- inviable), chc1∆ causes lethality. As a strategy to identify SCD1, we used pooled linkage analysis and whole genome sequencing. Here, we report that PAL2 (YHR097C) is the SCD1 locus. pal2∆ is synthetic lethal with chc1∆; whereas a deletion of its paralog, PAL1, is not synthetic lethal with clathrin deficiency. Like Pal1, Pal2 has two NPF motifs that are potential binding sites for EH domain proteins such as the early endocytic factor Ede1, and Pal2 associates with Ede1. Also, GFP-tagged Pal2p localizes to cortical patches containing other immobile phase endocytic coat factors. Overall, our data show that PAL2 is the SCD1 locus and the Pal2 protein has characteristics of an early factor involved in clathrin-mediated endocytosis. |
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Clathrin is a major coat protein involved in vesicle formation during endocytosis and transport in the endosomal/trans Golgi system. Clathrin is required for normal growth of yeast (Saccharomyces cerevisiae) and in some genetic backgrounds deletion of the clathrin heavy chain gene (CHC1) is lethal. Our lab defined a locus referred to as “suppressor of clathrin deficiency” (SCD1). In the presence of the scd1-v allele (“v” – viable), yeast cells lacking clathrin heavy chain survive but grow slowly, are morphologically abnormal and have many membrane trafficking defects. In the presence of scd1-i (“i”- inviable), chc1∆ causes lethality. As a strategy to identify SCD1, we used pooled linkage analysis and whole genome sequencing. Here, we report that PAL2 (YHR097C) is the SCD1 locus. pal2∆ is synthetic lethal with chc1∆; whereas a deletion of its paralog, PAL1, is not synthetic lethal with clathrin deficiency. Like Pal1, Pal2 has two NPF motifs that are potential binding sites for EH domain proteins such as the early endocytic factor Ede1, and Pal2 associates with Ede1. Also, GFP-tagged Pal2p localizes to cortical patches containing other immobile phase endocytic coat factors. Overall, our data show that PAL2 is the SCD1 locus and the Pal2 protein has characteristics of an early factor involved in clathrin-mediated endocytosis.</description><identifier>ISSN: 2160-1836</identifier><identifier>EISSN: 2160-1836</identifier><identifier>DOI: 10.1534/g3.118.200782</identifier><identifier>PMID: 30679249</identifier><language>eng</language><publisher>United States: Oxford University Press</publisher><subject>Clathrin ; Endocytosis ; Genetic Loci ; Investigations ; membrane trafficking ; pooled linkage analysis ; Receptors, Cell Surface - genetics ; Receptors, Cell Surface - metabolism ; Saccharomyces cerevisiae - metabolism ; Saccharomyces cerevisiae - physiology ; Saccharomyces cerevisiae Proteins - genetics ; Saccharomyces cerevisiae Proteins - metabolism ; Whole Genome Sequencing</subject><ispartof>G3 : genes - genomes - genetics, 2019-03, Vol.9 (3), p.867-877</ispartof><rights>2019 Moorthy et al. 2019</rights><rights>Copyright © 2019 Moorthy et al.</rights><rights>Copyright © 2019 Moorthy 2019</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c447t-6a0f2da97c7f0871388c817ce9b924658964fed95d263322cc6d72fd113c46163</citedby><cites>FETCH-LOGICAL-c447t-6a0f2da97c7f0871388c817ce9b924658964fed95d263322cc6d72fd113c46163</cites><orcidid>0000-0002-0591-443X</orcidid></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC6404604/pdf/$$EPDF$$P50$$Gpubmedcentral$$Hfree_for_read</linktopdf><linktohtml>$$Uhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC6404604/$$EHTML$$P50$$Gpubmedcentral$$Hfree_for_read</linktohtml><link.rule.ids>230,314,727,780,784,885,27924,27925,53791,53793</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/30679249$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Moorthy, Balaji T</creatorcontrib><creatorcontrib>Sharma, Anupam</creatorcontrib><creatorcontrib>Boettner, Douglas R</creatorcontrib><creatorcontrib>Wilson, Thomas E</creatorcontrib><creatorcontrib>Lemmon, Sandra K</creatorcontrib><title>Identification of Suppressor of Clathrin Deficiency-1 (SCD1) and Its Connection to Clathrin-Mediated Endocytosis in Saccharomyces cerevisiae</title><title>G3 : genes - genomes - genetics</title><addtitle>G3 (Bethesda)</addtitle><description>Abstract
Clathrin is a major coat protein involved in vesicle formation during endocytosis and transport in the endosomal/trans Golgi system. Clathrin is required for normal growth of yeast (Saccharomyces cerevisiae) and in some genetic backgrounds deletion of the clathrin heavy chain gene (CHC1) is lethal. Our lab defined a locus referred to as “suppressor of clathrin deficiency” (SCD1). In the presence of the scd1-v allele (“v” – viable), yeast cells lacking clathrin heavy chain survive but grow slowly, are morphologically abnormal and have many membrane trafficking defects. In the presence of scd1-i (“i”- inviable), chc1∆ causes lethality. As a strategy to identify SCD1, we used pooled linkage analysis and whole genome sequencing. Here, we report that PAL2 (YHR097C) is the SCD1 locus. pal2∆ is synthetic lethal with chc1∆; whereas a deletion of its paralog, PAL1, is not synthetic lethal with clathrin deficiency. Like Pal1, Pal2 has two NPF motifs that are potential binding sites for EH domain proteins such as the early endocytic factor Ede1, and Pal2 associates with Ede1. Also, GFP-tagged Pal2p localizes to cortical patches containing other immobile phase endocytic coat factors. Overall, our data show that PAL2 is the SCD1 locus and the Pal2 protein has characteristics of an early factor involved in clathrin-mediated endocytosis.</description><subject>Clathrin</subject><subject>Endocytosis</subject><subject>Genetic Loci</subject><subject>Investigations</subject><subject>membrane trafficking</subject><subject>pooled linkage analysis</subject><subject>Receptors, Cell Surface - genetics</subject><subject>Receptors, Cell Surface - metabolism</subject><subject>Saccharomyces cerevisiae - metabolism</subject><subject>Saccharomyces cerevisiae - physiology</subject><subject>Saccharomyces cerevisiae Proteins - genetics</subject><subject>Saccharomyces cerevisiae Proteins - metabolism</subject><subject>Whole Genome Sequencing</subject><issn>2160-1836</issn><issn>2160-1836</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2019</creationdate><recordtype>article</recordtype><sourceid>DOA</sourceid><recordid>eNqFkkFvFCEUxydGY5u1R6-GYz3MCgMDMxcTs626SY2H1TNh4LFLMwsjME32O_ihpd26tie5AI8_v_fg_avqLcFL0lL2YUuXhHTLBmPRNS-q84ZwXJOO8pdP1mfVRUq3uIy25Zzx19UZxVz0DevPq99rAz4767TKLngULNrM0xQhpRDvd6tR5V10Hl1BETnw-lATdLlZXZH3SHmD1jmhVfAe9AMgh9OV-hsYpzIYdO1N0IcckkuooDZK652KYX_QkJCGCHcuOQVvqldWjQkuHudF9fPz9Y_V1_rm-5f16tNNrRkTueYK28aoXmhhcScI7TrdEaGhH8qjeNv1nFkwfWsaTmnTaM2NaKwhhGrGCaeLan3kmqBu5RTdXsWDDMrJh0CIW6lidnoEKRjjhthWiUExjs1gMMGEGU2NhaHDhfXxyJrmYQ9Gl9-ManwGfX7i3U5uw53kDBcgK4DLR0AMv2ZIWe5d0jCOykOYk2yI6JkQbWn4oqqPUh1DShHsKQ3B8t4QcktlMYQ8GqLo3z2t7aT-2_5_ucM8_Yf1B2nevhw</recordid><startdate>20190301</startdate><enddate>20190301</enddate><creator>Moorthy, Balaji T</creator><creator>Sharma, Anupam</creator><creator>Boettner, Douglas R</creator><creator>Wilson, Thomas E</creator><creator>Lemmon, Sandra K</creator><general>Oxford University Press</general><general>Genetics Society of America</general><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>7X8</scope><scope>5PM</scope><scope>DOA</scope><orcidid>https://orcid.org/0000-0002-0591-443X</orcidid></search><sort><creationdate>20190301</creationdate><title>Identification of Suppressor of Clathrin Deficiency-1 (SCD1) and Its Connection to Clathrin-Mediated Endocytosis in Saccharomyces cerevisiae</title><author>Moorthy, Balaji T ; Sharma, Anupam ; Boettner, Douglas R ; Wilson, Thomas E ; Lemmon, Sandra K</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c447t-6a0f2da97c7f0871388c817ce9b924658964fed95d263322cc6d72fd113c46163</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2019</creationdate><topic>Clathrin</topic><topic>Endocytosis</topic><topic>Genetic Loci</topic><topic>Investigations</topic><topic>membrane trafficking</topic><topic>pooled linkage analysis</topic><topic>Receptors, Cell Surface - genetics</topic><topic>Receptors, Cell Surface - metabolism</topic><topic>Saccharomyces cerevisiae - metabolism</topic><topic>Saccharomyces cerevisiae - physiology</topic><topic>Saccharomyces cerevisiae Proteins - genetics</topic><topic>Saccharomyces cerevisiae Proteins - metabolism</topic><topic>Whole Genome Sequencing</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Moorthy, Balaji T</creatorcontrib><creatorcontrib>Sharma, Anupam</creatorcontrib><creatorcontrib>Boettner, Douglas R</creatorcontrib><creatorcontrib>Wilson, Thomas E</creatorcontrib><creatorcontrib>Lemmon, Sandra K</creatorcontrib><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>MEDLINE - Academic</collection><collection>PubMed Central (Full Participant titles)</collection><collection>Directory of Open Access Journals</collection><jtitle>G3 : genes - genomes - genetics</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Moorthy, Balaji T</au><au>Sharma, Anupam</au><au>Boettner, Douglas R</au><au>Wilson, Thomas E</au><au>Lemmon, Sandra K</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Identification of Suppressor of Clathrin Deficiency-1 (SCD1) and Its Connection to Clathrin-Mediated Endocytosis in Saccharomyces cerevisiae</atitle><jtitle>G3 : genes - genomes - genetics</jtitle><addtitle>G3 (Bethesda)</addtitle><date>2019-03-01</date><risdate>2019</risdate><volume>9</volume><issue>3</issue><spage>867</spage><epage>877</epage><pages>867-877</pages><issn>2160-1836</issn><eissn>2160-1836</eissn><abstract>Abstract
Clathrin is a major coat protein involved in vesicle formation during endocytosis and transport in the endosomal/trans Golgi system. Clathrin is required for normal growth of yeast (Saccharomyces cerevisiae) and in some genetic backgrounds deletion of the clathrin heavy chain gene (CHC1) is lethal. Our lab defined a locus referred to as “suppressor of clathrin deficiency” (SCD1). In the presence of the scd1-v allele (“v” – viable), yeast cells lacking clathrin heavy chain survive but grow slowly, are morphologically abnormal and have many membrane trafficking defects. In the presence of scd1-i (“i”- inviable), chc1∆ causes lethality. As a strategy to identify SCD1, we used pooled linkage analysis and whole genome sequencing. Here, we report that PAL2 (YHR097C) is the SCD1 locus. pal2∆ is synthetic lethal with chc1∆; whereas a deletion of its paralog, PAL1, is not synthetic lethal with clathrin deficiency. Like Pal1, Pal2 has two NPF motifs that are potential binding sites for EH domain proteins such as the early endocytic factor Ede1, and Pal2 associates with Ede1. Also, GFP-tagged Pal2p localizes to cortical patches containing other immobile phase endocytic coat factors. Overall, our data show that PAL2 is the SCD1 locus and the Pal2 protein has characteristics of an early factor involved in clathrin-mediated endocytosis.</abstract><cop>United States</cop><pub>Oxford University Press</pub><pmid>30679249</pmid><doi>10.1534/g3.118.200782</doi><tpages>11</tpages><orcidid>https://orcid.org/0000-0002-0591-443X</orcidid><oa>free_for_read</oa></addata></record> |
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subjects | Clathrin Endocytosis Genetic Loci Investigations membrane trafficking pooled linkage analysis Receptors, Cell Surface - genetics Receptors, Cell Surface - metabolism Saccharomyces cerevisiae - metabolism Saccharomyces cerevisiae - physiology Saccharomyces cerevisiae Proteins - genetics Saccharomyces cerevisiae Proteins - metabolism Whole Genome Sequencing |
title | Identification of Suppressor of Clathrin Deficiency-1 (SCD1) and Its Connection to Clathrin-Mediated Endocytosis in Saccharomyces cerevisiae |
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