title: Structure, dynamics and interactions of p47, a major adaptor of the AAA ATPase, p97.
abstract: p47 is a major adaptor molecule of the cytosolic AAA ATPase p97. The principal role of the p97-p47 complex is in regulation of membrane fusion events. Mono-ubiquitin recognition by p47 has also been shown to be crucial in the p97-p47-mediated Golgi membrane fusion events. Here, we describe the high-resolution solution structures of the N-terminal UBA domain and the central domain (SEP) from p47. The p47 UBA domain has the characteristic three-helix bundle fold and forms a highly stable complex with ubiquitin. We report the interaction surfaces of the two proteins and present a structure for the p47 UBA-ubiquitin complex. The p47 SEP domain adopts a novel fold with a betabetabetaalphaalphabeta secondary structure arrangement, where beta4 pairs in a parallel fashion to beta1. Based on biophysical studies, we demonstrate a clear propensity for the self-association of p47. Furthermore, p97 N binding abolishes p47 self-association, revealing the potential interaction surfaces for recognition of other domains within p97 or the substrate.
authors.affiliation: Department of Biological Sciences, Wolfson Laboratories, Imperial College London, South Kensington, London, UK.
authors.forename: Xuemei
authors.initials: X
authors.lastname: Yuan
authors.forename: Peter
authors.initials: P
authors.lastname: Simpson
authors.forename: Ciaran
authors.initials: C
authors.lastname: McKeown
authors.forename: Hisao
authors.initials: H
authors.lastname: Kondo
authors.forename: Keiji
authors.initials: K
authors.lastname: Uchiyama
authors.forename: Russell
authors.initials: R
authors.lastname: Wallis
authors.forename: Ingrid
authors.initials: I
authors.lastname: Dreveny
authors.forename: Catherine
authors.initials: C
authors.lastname: Keetch
authors.forename: Xiaodong
authors.initials: X
authors.lastname: Zhang
authors.forename: Carol
authors.initials: C
authors.lastname: Robinson
authors.forename: Paul
authors.initials: P
authors.lastname: Freemont
authors.forename: Stephen
authors.initials: S
authors.lastname: Matthews
mesh_terms: D048868:Adaptor Proteins, Signal Transducing / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D000251:Adenosine Triphosphatases / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D000595:Amino Acid Sequence; D000818:Animals; D006801:Humans; D008958:Models, Molecular; D008969:Molecular Sequence Data; D019906:Nuclear Magnetic Resonance, Biomolecular; D009687:Nuclear Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D017510:Protein Folding; D020836:Protein Structure, Quaternary; D017433:Protein Structure, Secondary; D051381:Rats; D016415:Sequence Alignment; D025801:Ubiquitin / Q000737:chemistry / Q000378:metabolism
chemical_list: D048868:Adaptor Proteins, Signal Transducing; D009687:Nuclear Proteins; D025801:Ubiquitin; D000251:Adenosine Triphosphatases; C404942:p97 ATPase