Identification of suppressor of Clathrin deficiency-1 (SCD1) and its connection to Clathrin-mediated endocytosis in Saccharomyces cerevisiae

Balaji T. Moorthy, Anupam Sharma, Douglas R. Boettner, Thomas E. Wilson, Sandra Lemmon

Research output: Contribution to journalArticle

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), chc1Δ 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. pal2Δ is synthetic lethal with chc1Δ; 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.

Original languageEnglish (US)
Pages (from-to)867-877
Number of pages11
JournalG3: Genes, Genomes, Genetics
Volume9
Issue number3
DOIs
StatePublished - Mar 1 2019

Fingerprint

Clathrin
Endocytosis
Saccharomyces cerevisiae
Clathrin Heavy Chains
Yeasts
Capsid Proteins
Alleles
Binding Sites
Genome
Membranes

Keywords

  • Clathrin
  • Endocytosis
  • Membrane trafficking
  • Pooled linkage analysis

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics
  • Genetics(clinical)

Cite this

Identification of suppressor of Clathrin deficiency-1 (SCD1) and its connection to Clathrin-mediated endocytosis in Saccharomyces cerevisiae. / Moorthy, Balaji T.; Sharma, Anupam; Boettner, Douglas R.; Wilson, Thomas E.; Lemmon, Sandra.

In: G3: Genes, Genomes, Genetics, Vol. 9, No. 3, 01.03.2019, p. 867-877.

Research output: Contribution to journalArticle

Moorthy, Balaji T. ; Sharma, Anupam ; Boettner, Douglas R. ; Wilson, Thomas E. ; Lemmon, Sandra. / Identification of suppressor of Clathrin deficiency-1 (SCD1) and its connection to Clathrin-mediated endocytosis in Saccharomyces cerevisiae. In: G3: Genes, Genomes, Genetics. 2019 ; Vol. 9, No. 3. pp. 867-877.
@article{bea4d89d0bf94166a5ef618e808811cf,
title = "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 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), chc1Δ 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. pal2Δ is synthetic lethal with chc1Δ; 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.",
keywords = "Clathrin, Endocytosis, Membrane trafficking, Pooled linkage analysis",
author = "Moorthy, {Balaji T.} and Anupam Sharma and Boettner, {Douglas R.} and Wilson, {Thomas E.} and Sandra Lemmon",
year = "2019",
month = "3",
day = "1",
doi = "10.1534/g3.118.200782",
language = "English (US)",
volume = "9",
pages = "867--877",
journal = "G3 (Bethesda, Md.)",
issn = "2160-1836",
publisher = "Genetics Society of America",
number = "3",

}

TY - JOUR

T1 - Identification of suppressor of Clathrin deficiency-1 (SCD1) and its connection to Clathrin-mediated endocytosis in Saccharomyces cerevisiae

AU - Moorthy, Balaji T.

AU - Sharma, Anupam

AU - Boettner, Douglas R.

AU - Wilson, Thomas E.

AU - Lemmon, Sandra

PY - 2019/3/1

Y1 - 2019/3/1

N2 - 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), chc1Δ 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. pal2Δ is synthetic lethal with chc1Δ; 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.

AB - 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), chc1Δ 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. pal2Δ is synthetic lethal with chc1Δ; 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.

KW - Clathrin

KW - Endocytosis

KW - Membrane trafficking

KW - Pooled linkage analysis

UR - http://www.scopus.com/inward/record.url?scp=85062623757&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=85062623757&partnerID=8YFLogxK

U2 - 10.1534/g3.118.200782

DO - 10.1534/g3.118.200782

M3 - Article

C2 - 30679249

AN - SCOPUS:85062623757

VL - 9

SP - 867

EP - 877

JO - G3 (Bethesda, Md.)

JF - G3 (Bethesda, Md.)

SN - 2160-1836

IS - 3

ER -