Structure, dynamics and interactions of p47, a major adaptor of the AAA ATPase, p97.
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.
Yuan
Xuemei
X
Department of Biological Sciences, Wolfson Laboratories, Imperial College London, South Kensington, London, UK.
Simpson
Peter
P
McKeown
Ciaran
C
Kondo
Hisao
H
Uchiyama
Keiji
K
Wallis
Russell
R
Dreveny
Ingrid
I
Keetch
Catherine
C
Zhang
Xiaodong
X
Robinson
Carol
C
Freemont
Paul
P
Matthews
Stephen
S
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
D048868:Adaptor Proteins, Signal Transducing; D009687:Nuclear Proteins; D025801:Ubiquitin; D000251:Adenosine Triphosphatases; C404942:p97 ATPase