Universität zu Köln

Rijal, Ramesh ORCID: 0000-0003-0498-1064, Arhzaouy, Khalid, Strucksberg, Karl-Heinz, Cross, Megan ORCID: 0000-0001-7512-7223, Hofmann, Andreas ORCID: 0000-0003-4408-5467, Schroeder, Rolf, Clemen, Christoph S. and Eichinger, Ludwig ORCID: 0000-0003-1594-6117 (2016). Mutant p97 exhibits species-specific changes of its ATPase activity and compromises the UBXD9-mediated monomerisation of p97 hexamers. Eur. J. Cell Biol., 95 (6-7). S. 195 - 208. MUNICH: ELSEVIER GMBH. ISSN 1618-1298

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Abstract

p97 (VCP) is a homo-hexameric triple-A ATPase that exerts a plethora of cellular processes. Heterozygous missense mutations of p97 cause at least five human neurodegenerative disorders. However, the specific molecular consequences of p97 mutations are hitherto widely unknown. Our in silico structural models of human and Dictyostelium p97 showed that the disease-causing human R93C, R155H, and R155C as well as Dictyostelium R154C, E219K, R154C/E219K p97 mutations constitute variations in surface-exposed locations. In-gel ATPase activity measurements of p97 monomers and hexamers revealed significant mutation- and species-specific differences. While all human p97 mutations led to an increase in ATPase activity, no changes could be detected for the Dictyostelium R154C mutant, which is orthologous to human R155C. The E219K mutation led to an almost complete loss of activity, which was partially recuperated in the R154C/E219K double-mutant indicating p97 inter-domain communication. By means of co-immunoprecipitation experiments we identified an UBX-domain containing Dictyostelium protein as a novel p97 interaction partner. We categorized all UBX-domain containing Dictyostelium proteins and named the interaction partner UBXD9. Pull-down assays and surface plasmon resonance analyses of Dictyostelium UBXD9 or the human orthologue TUG/ASPL/UBXD9 demonstrated direct interactions with p97 as well as species-, mutation- and ATP-dependent differences in the binding affinities. Sucrose density gradient assays revealed that both human and Dictyostelium UBXD9 proteins very efficiently disassembled wild-type, but to a lesser extent mutant p97 hexamers into monomers. Our results are consistent with a scenario in which p97 point mutations lead to differences in enzymatic activities and molecular interactions, which in the long-term result in a late-onset and progressive multisystem disease. (C) 2016 The Authors. Published by Elsevier GmbH.

Item Type:	Journal Article
Creators:	Creators Email ORCID ORCID Put Code Rijal, Ramesh UNSPECIFIED orcid.org/0000-0003-0498-1064 UNSPECIFIED Arhzaouy, Khalid UNSPECIFIED UNSPECIFIED UNSPECIFIED Strucksberg, Karl-Heinz UNSPECIFIED UNSPECIFIED UNSPECIFIED Cross, Megan UNSPECIFIED orcid.org/0000-0001-7512-7223 UNSPECIFIED Hofmann, Andreas UNSPECIFIED orcid.org/0000-0003-4408-5467 UNSPECIFIED Schroeder, Rolf UNSPECIFIED UNSPECIFIED UNSPECIFIED Clemen, Christoph S. UNSPECIFIED UNSPECIFIED UNSPECIFIED Eichinger, Ludwig UNSPECIFIED orcid.org/0000-0003-1594-6117 UNSPECIFIED
URN:	urn:nbn:de:hbz:38-273452
DOI:	10.1016/j.ejcb.2016.03.004
Journal or Publication Title:	Eur. J. Cell Biol.
Volume:	95
Number:	6-7
Page Range:	S. 195 - 208
Date:	2016
Publisher:	ELSEVIER GMBH
Place of Publication:	MUNICH
ISSN:	1618-1298
Language:	English
Faculty:	Unspecified
Divisions:	Unspecified
Subjects:	no entry
Uncontrolled Keywords:	Keywords Language AAA-ATPASE; UBX DOMAIN; DICTYOSTELIUM-DISCOIDEUM; CONFORMATIONAL-CHANGES; P97/VCP ATPASE; VCP MUTATIONS; PAGET-DISEASE; N-DOMAIN; PROTEIN; D1 Multiple languages Cell Biology Multiple languages
Refereed:	Yes
URI:	http://kups.ub.uni-koeln.de/id/eprint/27345