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
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Research Support, Non-U.S. Gov't
MeSH terms
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DNA-Binding Proteins*
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Fluorescent Antibody Technique
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Fungal Proteins / genetics
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Fungal Proteins / metabolism*
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In Situ Hybridization
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Myosin Heavy Chains*
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Myosin Type V*
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Myosins / metabolism*
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Nucleic Acid Conformation
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Precipitin Tests
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Protein Binding
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RNA, Fungal / chemistry
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RNA, Fungal / genetics
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RNA, Fungal / metabolism*
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RNA, Messenger / chemistry
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RNA, Messenger / genetics
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RNA, Messenger / metabolism*
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RNA-Binding Proteins / genetics
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RNA-Binding Proteins / metabolism*
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Recombinant Fusion Proteins / metabolism
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Repressor Proteins*
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Saccharomyces cerevisiae / cytology
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Saccharomyces cerevisiae / genetics
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Saccharomyces cerevisiae / metabolism*
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Saccharomyces cerevisiae Proteins*
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Transcription Factors / genetics*
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Two-Hybrid System Techniques
Substances
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ASH1 protein, S cerevisiae
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DNA-Binding Proteins
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Fungal Proteins
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Myo4 protein, S cerevisiae
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RNA, Fungal
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RNA, Messenger
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RNA-Binding Proteins
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Recombinant Fusion Proteins
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Repressor Proteins
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Saccharomyces cerevisiae Proteins
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Transcription Factors
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Myosin Type V
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Myosin Heavy Chains
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Myosins