Isolation of Saccharomyces cerevisiae RNase T1 hypersensitive (rns) mutants and genetic analysis of the RNS1/DSL1 gene

Tomoki Ishikawa, Kenji Unno, Gen Nonaka, Harushi Nakajima, Katsuhiko Kitamoto*

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

3 Scopus citations

Abstract

Overexpression of the rntA cDNA encoding RNase T1 derived from A. oryzae causes severe growth inhibition in S. cerevisiae. We previously reported that most S. cerevisiae mutant strains defective in translocation into the ER, ER-Golgi transport and vacuole formation exhibited hypersensitivity to expression of RNase T1. Screening for S. cerevisiae mutants that showed RNase T1 hypersensitivity resulted in the isolation of 38 (rns) mutant strains. Some of these mutants showed a variety of phenotypes including temperature-sensitive growth, hypersensitivity to G418, defect in invertase glycosylation and fragmented vacuoles. We identified the genes mutated in three of the rns mutants, rns1, rns2, and rns3, as DSL1, UMP1, and SEC17, respectively. Fluorescence microscopic observation showed that GFP or myc-tagged Rns1p was localized at the nuclear region in the cell. Two-hybrid screening revealed the interaction of Rns1p with a transcription factor Cin5p and a functionally unknown Ylr440cp. It was observed that HA-tagged Ylr440cp was localized to the ER and nuclear envelope.

Original languageEnglish (US)
Pages (from-to)73-82
Number of pages10
JournalJournal of General and Applied Microbiology
Volume51
Issue number2
DOIs
StatePublished - 2005
Externally publishedYes

Keywords

  • Proteasome
  • RNS1/DSL1
  • RNase T1
  • Saccharomyces cerevisiae
  • YLR440c

ASJC Scopus subject areas

  • Microbiology
  • Applied Microbiology and Biotechnology

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