She2p is a novel RNA-binding protein that recruits the Myo4p-She3p complex to ASH1 mRNA.
In Saccharomyces cerevisiae, Ash1p is a specific repressor of transcription that localizes exclusively to daughter cell nuclei through the asymmetric localization of ASH1 mRNA. This localization requires four cis-acting localization elements located in the ASH1 mRNA, five trans-acting factors, one of which is a myosin, and the actin cytoskeleton. The RNA-binding proteins that interact with these cis-elements remained to be identified. Starting with the 3' most localization element of ASH1 mRNA in the three-hybrid assay, element E3, we isolated a clone corresponding to the C-terminus of She3p. We also found that She3p and She2p interact, and this interaction is essential for the binding of She3p with element E3 in vivo. Moreover, She2p was observed to bind the E3 RNA directly in vitro and each of the ASH1 cis-acting localization elements requires She2p for their localization function. By tethering a She3p-MS2 fusion protein to a reporter RNA containing MS2 binding sites, we observed that She2p is dispensable for She3p-MS2-dependent RNA localization.
Long
R M
RM
Medical College of Wisconsin, Department of Microbiology and Molecular Genetics, 8701 Watertown Plank Road, Milwaukee, WI 53226-0509, USA. rlong@mcw.edu
Gu
W
W
Lorimer
E
E
Singer
R H
RH
Chartrand
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D000199:Actins / Q000378:metabolism; D002467:Cell Nucleus; D003599:Cytoskeleton / Q000378:metabolism; D004268:DNA-Binding Proteins; D004926:Escherichia coli / Q000378:metabolism; D005656:Fungal Proteins / Q000378:metabolism*; D017403:In Situ Hybridization; D007763:Lac Operon; D008954:Models, Biological; D018995:Myosin Heavy Chains; D024701:Myosin Type V; D009218:Myosins / Q000378:metabolism*; D010957:Plasmids / Q000378:metabolism; D011485:Protein Binding; D012333:RNA, Messenger / Q000378:metabolism*; D016601:RNA-Binding Proteins / Q000737:chemistry / Q000378:metabolism / Q000502:physiology*; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D012097:Repressor Proteins; D012441:Saccharomyces cerevisiae / Q000378:metabolism; D029701:Saccharomyces cerevisiae Proteins; D014157:Transcription Factors / Q000378:metabolism*; D014158:Transcription, Genetic; D015533:Transcriptional Activation; D020798:Two-Hybrid System Techniques; D014466:Ultraviolet Rays
C098733:ASH1 protein, S cerevisiae; D000199:Actins; D004268:DNA-Binding Proteins; D005656:Fungal Proteins; C098732:Myo4 protein, S cerevisiae; D012333:RNA, Messenger; D016601:RNA-Binding Proteins; D011993:Recombinant Fusion Proteins; D012097:Repressor Proteins; D029701:Saccharomyces cerevisiae Proteins; D014157:Transcription Factors; D024701:Myosin Type V; D018995:Myosin Heavy Chains; D009218:Myosins