She2p, a novel RNA-binding protein tethers ASH1 mRNA to the Myo4p myosin motor via She3p

EMBO J. 2000 Oct 16;19(20):5514-24. doi: 10.1093/emboj/19.20.5514.

Abstract

RNA localization is a widespread mechanism to achieve localized protein synthesis. In budding yeast, localization of ASH1 mRNA controls daughter cell-specific accumulation of the transcriptional regulator Ash1p, which determines mating type switching. ASH1 mRNA localization depends on four independently acting sequences ('zipcodes') within the mRNA. In addition, the class V myosin Myo4p and a set of She proteins with as yet unknown function are essential for ASH1 localization. Here we show that She2p is a novel RNA-binding protein that binds specifically to ASH1 mRNA in vivo and to ASH1 RNA zip codes in vitro. She2p can interact with She3 protein via She3p's C-terminus and becomes localized to the daughter cell tip upon ASH1 expression. The N-terminal coiled-coil domain of She3p is required to form an RNA-independent complex with the heavy chain of the myosin motor protein Myo4p. She2p and She3p are the first examples of adapters for tethering a localized mRNA to the motor protein and might serve as prototypes for RNA-motor protein adapters.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • DNA-Binding Proteins*
  • Fluorescent Antibody Technique
  • Fungal Proteins / genetics
  • Fungal Proteins / metabolism*
  • In Situ Hybridization
  • Myosin Heavy Chains*
  • Myosin Type V*
  • Myosins / metabolism*
  • Nucleic Acid Conformation
  • Precipitin Tests
  • Protein Binding
  • RNA, Fungal / chemistry
  • RNA, Fungal / genetics
  • RNA, Fungal / metabolism*
  • RNA, Messenger / chemistry
  • RNA, Messenger / genetics
  • RNA, Messenger / metabolism*
  • RNA-Binding Proteins / genetics
  • RNA-Binding Proteins / metabolism*
  • Recombinant Fusion Proteins / metabolism
  • Repressor Proteins*
  • Saccharomyces cerevisiae / cytology
  • Saccharomyces cerevisiae / genetics
  • Saccharomyces cerevisiae / metabolism*
  • Saccharomyces cerevisiae Proteins*
  • Transcription Factors / genetics*
  • Two-Hybrid System Techniques

Substances

  • ASH1 protein, S cerevisiae
  • DNA-Binding Proteins
  • Fungal Proteins
  • Myo4 protein, S cerevisiae
  • RNA, Fungal
  • RNA, Messenger
  • RNA-Binding Proteins
  • Recombinant Fusion Proteins
  • Repressor Proteins
  • Saccharomyces cerevisiae Proteins
  • Transcription Factors
  • Myosin Type V
  • Myosin Heavy Chains
  • Myosins