Abstract
Puf3p binds preferentially to messenger RNAs (mRNAs) for nuclear-encoded mitochondrial proteins. We find that Puf3p localizes to the cytosolic face of the mitochondrial outer membrane. Overexpression of PUF3 results in reduced mitochondrial respiratory activity and reduced levels of Pet123p, a protein encoded by a Puf3p-binding mRNA. Puf3p levels are reduced during the diauxic shift and growth on a nonfermentable carbon source, conditions that stimulate mitochondrial biogenesis. These findings support a role for Puf3p in mitochondrial biogenesis through effects on mRNA interactions. In addition, Puf3p links the mitochore, a complex required for mitochondrial-cytoskeletal interactions, to the Arp2/3 complex, the force generator for actin-dependent, bud-directed mitochondrial movement. Puf3p, the mitochore, and the Arp2/3 complex coimmunoprecipitate and have two-hybrid interactions. Moreover, deletion of PUF3 results in reduced interaction between the mitochore and the Arp2/3 complex and defects in mitochondrial morphology and motility similar to those observed in Arp2/3 complex mutants. Thus, Puf3p is a mitochondrial protein that contributes to the biogenesis and motility of the organelle.
Publication types
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Research Support, N.I.H., Extramural
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Research Support, Non-U.S. Gov't
MeSH terms
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Actin-Related Protein 2-3 Complex / genetics
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Actin-Related Protein 2-3 Complex / metabolism
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Binding Sites / genetics
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Cell Division / genetics
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Cell Division / physiology
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Immunoprecipitation
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Membrane Proteins / genetics
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Membrane Proteins / metabolism
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Membrane Transport Proteins / genetics
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Membrane Transport Proteins / metabolism
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Mitochondria / metabolism*
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Mitochondria / physiology
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Mitochondrial Membrane Transport Proteins
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Mitochondrial Membranes / metabolism
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Mitochondrial Precursor Protein Import Complex Proteins
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Mitochondrial Proteins / genetics
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Mitochondrial Proteins / metabolism
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Mutation
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Protein Binding / genetics
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RNA-Binding Proteins / genetics
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RNA-Binding Proteins / metabolism*
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RNA-Binding Proteins / physiology
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Ribosomal Proteins / genetics
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Ribosomal Proteins / metabolism
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Saccharomyces cerevisiae / genetics
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Saccharomyces cerevisiae / metabolism*
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Saccharomyces cerevisiae / physiology
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Saccharomyces cerevisiae Proteins / genetics
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Saccharomyces cerevisiae Proteins / metabolism*
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Saccharomyces cerevisiae Proteins / physiology
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Transformation, Genetic
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Two-Hybrid System Techniques
Substances
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Actin-Related Protein 2-3 Complex
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Membrane Proteins
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Membrane Transport Proteins
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Mitochondrial Membrane Transport Proteins
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Mitochondrial Precursor Protein Import Complex Proteins
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Mitochondrial Proteins
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PET123 protein, S cerevisiae
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PUF3 protein, S cerevisiae
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RNA-Binding Proteins
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Ribosomal Proteins
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SAM37 protein, S cerevisiae
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Saccharomyces cerevisiae Proteins
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TOM7 protein, S cerevisiae