Universität zu Köln

Arhzaouy, Khalid, Strucksberg, Karl-Heinz, Tung, Sze Man, Tangavelou, Karthikeyan, Stumpf, Maria, Faix, Jan, Schroeder, Rolf, Clemen, Christoph S. and Eichinger, Ludwig ORCID: 0000-0003-1594-6117 (2012). Heteromeric p97/p97(R155C) Complexes Induce Dominant Negative Changes in Wild- Type and Autophagy 9-Deficient Dictyostelium strains. PLoS One, 7 (10). SAN FRANCISCO: PUBLIC LIBRARY SCIENCE. ISSN 1932-6203

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Abstract

Heterozygous mutations in the human VCP (p97) gene cause autosomal-dominant IBMPFD (inclusion body myopathy with early onset Paget's disease of bone and frontotemporal dementia), ALS14 (amyotrophic lateral sclerosis with or without frontotemporal dementia) and HSP (hereditary spastic paraplegia). Most prevalent is the R155C point mutation. We studied the function of p97 in the social amoeba Dictyostelium discoideum and have generated strains that ectopically express wildtype (p97) or mutant p97 (p97(R155C)) fused to RFP in AX2 wild-type and autophagy 9 knock-out (ATG9(KO)) cells. Native gel electrophoresis showed that both p97 and p97(R155C) assemble into hexamers. Co-immunoprecipitation studies revealed that endogenous p97 and p97(R155C)-RFP form heteromers. The mutant strains displayed changes in cell growth, phototaxis, development, proteasomal activity, ubiquitinylated proteins, and ATG8(LC3) indicating mis-regulation of multiple essential cellular processes. Additionally, immunofluorescence analysis revealed an increase of protein aggregates in ATG9(KO)/p97(R155C)-RFP and ATG9(KO) cells. They were positive for ubiquitin in both strains, however, solely immunoreactive for p97 in the ATG9(KO) mutant. A major finding is that the expression of p97(R155C)-RFP in the ATG9(KO) strain partially or fully rescued the pleiotropic phenotype. We also observed dose-dependent effects of p97 on several cellular processes. Based on findings in the single versus the double mutants we propose a novel mode of p97 interaction with the core autophagy protein ATG9 which is based on mutual inhibition.

Item Type:	Journal Article
Creators:	Creators Email ORCID ORCID Put Code Arhzaouy, Khalid UNSPECIFIED UNSPECIFIED UNSPECIFIED Strucksberg, Karl-Heinz UNSPECIFIED UNSPECIFIED UNSPECIFIED Tung, Sze Man UNSPECIFIED UNSPECIFIED UNSPECIFIED Tangavelou, Karthikeyan UNSPECIFIED UNSPECIFIED UNSPECIFIED Stumpf, Maria UNSPECIFIED UNSPECIFIED UNSPECIFIED Faix, Jan UNSPECIFIED UNSPECIFIED 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:38-481165
DOI:	10.1371/journal.pone.0046879
Journal or Publication Title:	PLoS One
Volume:	7
Number:	10
Date:	2012
Publisher:	PUBLIC LIBRARY SCIENCE
Place of Publication:	SAN FRANCISCO
ISSN:	1932-6203
Language:	English
Faculty:	Unspecified
Divisions:	Unspecified
Subjects:	no entry
Uncontrolled Keywords:	Keywords Language VALOSIN-CONTAINING PROTEIN; INCLUSION-BODY MYOPATHY; PAGET-DISEASE; AAA-ATPASE; VCP; P97; DISCOIDEUM; CLEARANCE; MUTATIONS; BONE Multiple languages Multidisciplinary Sciences Multiple languages
URI:	http://kups.ub.uni-koeln.de/id/eprint/48116