Abstract
Get3, Get4, and Get5 in Saccharomyces cerevisiae participate in the insertion of tail-anchored proteins into the endoplasmic reticulum membrane. We elucidated the interaction between Get4 and Get5 and investigated their interaction with Get3 and a tetratricopeptide repeat-containing protein, Sgt2. Based on co-immunoprecipitation and crystallographic studies, Get4 and Get5 formed a tight complex, suggesting that they constitute subunits of a larger complex. In contrast, although Get3 interacted physically with the Get4-Get5 complex, low amounts of Get3 co-precipitated with Get5, implying a transient interaction between Get3 and Get4-Get5. Sgt2 also interacted with Get5, although the amount of Sgt2 that co-precipitated with Get5 varied. Moreover, GET3, GET4, and GET5 interacted genetically with molecular chaperone YDJ1, suggesting that chaperones might also be involved in the insertion of tail-anchored proteins.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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Adenosine Triphosphatases / metabolism*
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Carrier Proteins / chemistry*
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Carrier Proteins / metabolism*
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Crystallography, X-Ray / methods
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Fungal Proteins / chemistry
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Fungal Proteins / metabolism*
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Gene Deletion
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Gene Expression Profiling
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Gene Expression Regulation, Fungal*
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Guanine Nucleotide Exchange Factors / metabolism*
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HSP40 Heat-Shock Proteins / metabolism*
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Mass Spectrometry / methods
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Membrane Proteins
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Molecular Chaperones / metabolism
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Protein Interaction Mapping
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Protein Structure, Tertiary
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Protein Transport
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Saccharomyces cerevisiae / enzymology*
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Saccharomyces cerevisiae Proteins / chemistry*
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Saccharomyces cerevisiae Proteins / metabolism*
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Two-Hybrid System Techniques
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Ubiquitin / chemistry*
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Ubiquitin / metabolism
Substances
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Carrier Proteins
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Fungal Proteins
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GET4 protein, S cerevisiae
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Guanine Nucleotide Exchange Factors
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HSP40 Heat-Shock Proteins
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Mdy2 protein, S cerevisiae
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Membrane Proteins
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Molecular Chaperones
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Saccharomyces cerevisiae Proteins
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Sgt2 protein, S cerevisiae
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Ubiquitin
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YDJ1 protein, S cerevisiae
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Adenosine Triphosphatases
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Get3 protein, S cerevisiae