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 K. 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

Keywords

  • Clathrin
  • Endocytosis
  • Membrane trafficking
  • Pooled linkage analysis

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics
  • Genetics(clinical)

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