Parallel analysis of tagged deletion mutants efficiently identifies genes involved in endoplasmic reticulum biogenesis

Robin Wright, Mark L. Parrish, Emily Cadera, Lynnelle Larson, Clinton K. Matson, Philip Garrett-Engele, Chris Armour, Pek Yee Lum, Daniel D. Shoemaker

Research output: Contribution to journalArticlepeer-review

23 Scopus citations

Abstract

Increased levels of HMG-CoA reductase induce cell type- and isozyme-specific proliferation of the endoplasmic reticulum. In yeast, the ER proliferations induced by Hmg1p consist of nuclear-associated stacks of smooth ER membranes known as karmellae. To identify genes required for karmellae assembly, we compared the composition of populations of homozygous diploid S. cerevisiae deletion mutants following 20 generations of growth with and without karmellae. Using an initial population of 1557 deletion mutants, 120 potential mutants were identified as a result of three independent experiments. Each experiment produced a largely nonoverlapping set of potential mutants, suggesting that differences in specific growth conditions could be used to maximize the comprehensiveness of similar parallel analysis screens. Only two genes, UBC7 and YAL011W, were identified in all three experiments. Subsequent analysis of individual mutant strains confirmed that each experiment was identifying valid mutations, based on the mutant's sensitivity to elevated HMG-CoA reductase and inability to assemble normal karmellae. The largest class of HMG-CoA reductase-sensitive mutations was a subset of genes that are involved in chromatin structure and transcriptional regulation, suggesting that karmellae assembly requires changes in transcription or that the presence of karmellae may interfere with normal transcriptional regulation.

Original languageEnglish (US)
Pages (from-to)881-892
Number of pages12
JournalYeast
Volume20
Issue number10
DOIs
StatePublished - Jul 1 2003

Keywords

  • DNA microarray
  • Endoplasmic reticulum
  • HMG-CoA reductase
  • Karmellae
  • Saccharomyces cerevisiae

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