title: NMR studies on the interactions between yeast Vta1 and Did2 during the multivesicular bodies sorting pathway.
abstract: As an AAA-ATPase, Vps4 is important for function of multivesicular bodies (MVB) sorting pathway, which involves in cellular phenomena ranging from receptor down-regulation to viral budding to cytokinesis. The activity of Vps4 is stimulated by the interactions between Vta1 N-terminus (named as Vta1NTD) and Did2 fragment (176-204 aa) (termed as Did2176-204) or Vps60 (128-186 aa) (termed as Vps60128-186). The structural basis of how Vta1NTD binds to Did2176-204is still unclear. To address this, in this report, the structure of Did2176-204in complex with Vta1NTD was determined by NMR techniques, demonstrating that Did2176-204interacts with Vta1NTD through its helix α6' extending over the 2ndand the 3rdα-helices of Vta1NTD microtubule interacting and transport 1 (MIT1) domain. The residues within Did2176-204helix α6' in the interface make up of an amino acid sequence as E192'xxL195'xxR198'L199'xxL202'R203', identical to type 1 MIT-interacting motif (MIM1) (D/E)xxLxxRLxxL(K/R) of CHMP1A180-196observed in Vps4-CHMP1A complex structure, indicating that Did2 binds to Vta1NTD through canonical MIM1 interactions. Moreover, the Did2 binding does not result in Vta1NTD significant conformational changes, revealing that Did2, similar to Vps60, enhances Vta1 stimulation of Vps4 ATPase activity in an indirect manner.
authors.affiliation: State Key Laboratory of Bioorganic and Natural Products Chemistry and Collaborative Innovation Center of Chemistry for Life Sciences, Shanghai Institute of Organic Chemistry, Chinese Academy of Sciences, 345 Lingling Road, Shanghai, 200032, China.|Tianjin Institute of Industrial Biotechnology, Chinese Academy of Sciences, 32 XiQiDao, Tianjin Airport Economic Area, Tianjin, 300308, China.
authors.forename: Jie
authors.initials: J
authors.lastname: Shen
authors.affiliation: State Key Laboratory of Bioorganic and Natural Products Chemistry and Collaborative Innovation Center of Chemistry for Life Sciences, Shanghai Institute of Organic Chemistry, Chinese Academy of Sciences, 345 Lingling Road, Shanghai, 200032, China.|Public Technology Service Platform, Wuhan Institute of Biotechnology, 666 Gaoxin Road, Wuhan, 430075, China.
authors.forename: Zhongzheng
authors.initials: Z
authors.lastname: Yang
authors.affiliation: State Key Laboratory of Bioorganic and Natural Products Chemistry and Collaborative Innovation Center of Chemistry for Life Sciences, Shanghai Institute of Organic Chemistry, Chinese Academy of Sciences, 345 Lingling Road, Shanghai, 200032, China.
authors.forename: Jiaolong
authors.initials: J
authors.lastname: Wang
authors.affiliation: State Key Laboratory of Bioorganic and Natural Products Chemistry and Collaborative Innovation Center of Chemistry for Life Sciences, Shanghai Institute of Organic Chemistry, Chinese Academy of Sciences, 345 Lingling Road, Shanghai, 200032, China.
authors.forename: Bin
authors.initials: B
authors.lastname: Zhao
authors.affiliation: State Key Laboratory of Bioorganic and Natural Products Chemistry and Collaborative Innovation Center of Chemistry for Life Sciences, Shanghai Institute of Organic Chemistry, Chinese Academy of Sciences, 345 Lingling Road, Shanghai, 200032, China.
authors.forename: Wenxian
authors.initials: W
authors.lastname: Lan
authors.affiliation: State Key Laboratory of Bioorganic and Natural Products Chemistry and Collaborative Innovation Center of Chemistry for Life Sciences, Shanghai Institute of Organic Chemistry, Chinese Academy of Sciences, 345 Lingling Road, Shanghai, 200032, China.
authors.forename: Chunxi
authors.initials: C
authors.lastname: Wang
authors.affiliation: State Key Laboratory of Magnetic Resonance and Atomic and Molecular Physics, Wuhan Institute of Physics and Mathematics, Chinese Academy of Sciences, 30 West Xiaohongshan Road, Wuhan, 430071, China.
authors.forename: Xu
authors.initials: X
authors.lastname: Zhang
authors.affiliation: Life Sciences Institute and Department of Biological Chemistry, Medical School, University of Michigan, 210 Washtenaw Ave, Ann Arbor, MI 48109, USA.
authors.forename: Cody J
authors.initials: CJ
authors.lastname: Wild
authors.affiliation: State Key Laboratory of Magnetic Resonance and Atomic and Molecular Physics, Wuhan Institute of Physics and Mathematics, Chinese Academy of Sciences, 30 West Xiaohongshan Road, Wuhan, 430071, China.
authors.forename: Maili
authors.initials: M
authors.lastname: Liu
authors.affiliation: Life Sciences Institute and Department of Biological Chemistry, Medical School, University of Michigan, 210 Washtenaw Ave, Ann Arbor, MI 48109, USA.
authors.forename: Zhaohui
authors.initials: Z
authors.lastname: Xu
authors.affiliation: State Key Laboratory of Bioorganic and Natural Products Chemistry and Collaborative Innovation Center of Chemistry for Life Sciences, Shanghai Institute of Organic Chemistry, Chinese Academy of Sciences, 345 Lingling Road, Shanghai, 200032, China.
authors.forename: Chunyang
authors.initials: C
authors.lastname: Cao
mesh_terms: D020816:Amino Acid Motifs; D056827:Endosomal Sorting Complexes Required for Transport / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D019906:Nuclear Magnetic Resonance, Biomolecular; D011485:Protein Binding; D000072417:Protein Domains; D012441:Saccharomyces cerevisiae / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D029701:Saccharomyces cerevisiae Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism
chemical_list: C511158:DID2 protein, S cerevisiae; D056827:Endosomal Sorting Complexes Required for Transport; D029701:Saccharomyces cerevisiae Proteins; C487285:VTA1 protein, S cerevisiae