VCP binding influences intracellular distribution of the slow Wallerian degeneration protein, Wld(S).
Wallerian degeneration slow (Wld(S)) mice express a chimeric protein that delays axonal degeneration. The N-terminal domain (N70), which is essential for axonal protection in vivo, binds valosin-containing protein (VCP) and targets both Wld(S) and VCP to discrete nuclear foci. We characterized the formation, composition and localization of these potentially important foci. Missense mutations show that the N-terminal sixteen residues (N16) of Wld(S) are essential for both VCP binding and targeting Wld(S) to nuclear foci. Removing N16 abolishes foci, and VCP binding sequences from ataxin-3 or HrdI restore them. In vitro, these puncta co-localize with proteasome subunits. In vivo, Wld(S) assumes a range of nuclear distribution patterns, including puncta, and its neuronal expression and intranuclear distribution is region-specific and varies between spontaneous and transgenic Wld(S) models. We conclude that VCP influences Wld(S) intracellular distribution, and thus potentially its function, by binding within the N70 domain required for axon protection.
Wilbrey
Anna L
AL
The Babraham Institute, Babraham Research Campus, Cambridge, CB22 3AT, UK.
Haley
Jane E
JE
Wishart
Thomas M
TM
Conforti
Laura
L
Morreale
Giacomo
G
Beirowski
Bogdan
B
Babetto
Elisabetta
E
Adalbert
Robert
R
Gillingwater
Thomas H
TH
Smith
Trevor
T
Wyllie
David J A
DJ
Ribchester
Richard R
RR
Coleman
Michael P
MP
D000251:Adenosine Triphosphatases / Q000032:analysis / Q000235:genetics / Q000378:metabolism*; D000818:Animals; D030801:Animals, Genetically Modified; D001923:Brain Chemistry / Q000235:genetics; D018797:Cell Cycle Proteins / Q000032:analysis / Q000235:genetics / Q000378:metabolism*; D002478:Cells, Cultured; D003593:Cytoplasm / Q000737:chemistry / Q000378:metabolism; D006367:HeLa Cells; D006801:Humans; D007424:Intracellular Fluid / Q000737:chemistry / Q000378:metabolism*; D051379:Mice; D008810:Mice, Inbred C57BL; D008822:Mice, Transgenic; D020125:Mutation, Missense; D009419:Nerve Tissue Proteins / Q000032:analysis / Q000235:genetics / Q000378:metabolism*; D016716:PC12 Cells; D011485:Protein Binding / Q000502:physiology; D051381:Rats; D000074405:Valosin Containing Protein; D014855:Wallerian Degeneration / Q000235:genetics / Q000378:metabolism
D018797:Cell Cycle Proteins; D009419:Nerve Tissue Proteins; C442580:Wld protein, mouse; D000251:Adenosine Triphosphatases; C000614178:VCP protein, human; D000074405:Valosin Containing Protein; C000614179:Vcp protein, mouse; C000614180:Vcp protein, rat