Crystal structure of Get4-Get5 complex and its interactions with Sgt2, Get3, and Ydj1.
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.
Chang
Yi-Wei
YW
Institute of Molecular Biology, Academia Sinica, Taipei 115; Institute of Bioinformatics and Structural Biology, National Tsing Hua University, Hsinchu 300.
Chuang
Yu-Chien
YC
Institute of Molecular Biology, Academia Sinica, Taipei 115.
Ho
Yu-Chi
YC
Institute of Molecular Biology, Academia Sinica, Taipei 115.
Cheng
Ming-Yuan
MY
Institute of Genome Sciences, National Yang-Ming University, Taipei 112, Taiwan.
Sun
Yuh-Ju
YJ
Institute of Bioinformatics and Structural Biology, National Tsing Hua University, Hsinchu 300.
Hsiao
Chwan-Deng
CD
Institute of Molecular Biology, Academia Sinica, Taipei 115. Electronic address: hsiao@gate.sinica.edu.tw.
Wang
Chung
C
Institute of Molecular Biology, Academia Sinica, Taipei 115. Electronic address: mbcw@gate.sinica.edu.tw.
D000251:Adenosine Triphosphatases / Q000378:metabolism*; D002352:Carrier Proteins / Q000737:chemistry* / Q000378:metabolism*; D018360:Crystallography, X-Ray / Q000379:methods; D005656:Fungal Proteins / Q000737:chemistry / Q000378:metabolism*; D017353:Gene Deletion; D020869:Gene Expression Profiling; D015966:Gene Expression Regulation, Fungal; D020662:Guanine Nucleotide Exchange Factors / Q000378:metabolism*; D050956:HSP40 Heat-Shock Proteins / Q000378:metabolism*; D013058:Mass Spectrometry / Q000379:methods; D008565:Membrane Proteins; D018832:Molecular Chaperones / Q000378:metabolism; D025941:Protein Interaction Mapping; D017434:Protein Structure, Tertiary; D021381:Protein Transport; D012441:Saccharomyces cerevisiae / Q000201:enzymology*; D029701:Saccharomyces cerevisiae Proteins / Q000737:chemistry* / Q000378:metabolism*; D020798:Two-Hybrid System Techniques; D025801:Ubiquitin / Q000737:chemistry* / Q000378:metabolism
D002352:Carrier Proteins; D005656:Fungal Proteins; C548889:GET4 protein, S cerevisiae; D020662:Guanine Nucleotide Exchange Factors; D050956:HSP40 Heat-Shock Proteins; C510068:Mdy2 protein, S cerevisiae; D008565:Membrane Proteins; D018832:Molecular Chaperones; D029701:Saccharomyces cerevisiae Proteins; C520576:Sgt2 protein, S cerevisiae; D025801:Ubiquitin; C070319:YDJ1 protein, S cerevisiae; D000251:Adenosine Triphosphatases; C529048:Get3 protein, S cerevisiae