title: The slow Wallerian degeneration protein, WldS, binds directly to VCP/p97 and partially redistributes it within the nucleus.
abstract: Slow Wallerian degeneration (Wld(S)) mutant mice express a chimeric nuclear protein that protects sick or injured axons from degeneration. The C-terminal region, derived from NAD(+) synthesizing enzyme Nmnat1, is reported to confer neuroprotection in vitro. However, an additional role for the N-terminal 70 amino acids (N70), derived from multiubiquitination factor Ube4b, has not been excluded. In wild-type Ube4b, N70 is part of a sequence essential for ubiquitination activity but its role is not understood. We report direct binding of N70 to valosin-containing protein (VCP; p97/Cdc48), a protein with diverse cellular roles including a pivotal role in the ubiquitin proteasome system. Interaction with Wld(S) targets VCP to discrete intranuclear foci where ubiquitin epitopes can also accumulate. Wld(S) lacking its N-terminal 16 amino acids (N16) neither binds nor redistributes VCP, but continues to accumulate in intranuclear foci, targeting its intrinsic NAD(+) synthesis activity to these same foci. Wild-type Ube4b also requires N16 to bind VCP, despite a more C-terminal binding site in invertebrate orthologues. We conclude that N-terminal sequences of Wld(S) protein influence the intranuclear location of both ubiquitin proteasome and NAD(+) synthesis machinery and that an evolutionary recent sequence mediates binding of mammalian Ube4b to VCP.
authors.affiliation: Institute for Genetics and Center for Molecular Medicine, University of Cologne, D-50674 Cologne, Germany.
authors.forename: Heike
authors.initials: H
authors.lastname: Laser
authors.forename: Laura
authors.initials: L
authors.lastname: Conforti
authors.forename: Giacomo
authors.initials: G
authors.lastname: Morreale
authors.forename: Till G M
authors.initials: TG
authors.lastname: Mack
authors.forename: Molly
authors.initials: M
authors.lastname: Heyer
authors.forename: Jane E
authors.initials: JE
authors.lastname: Haley
authors.forename: Thomas M
authors.initials: TM
authors.lastname: Wishart
authors.forename: Bogdan
authors.initials: B
authors.lastname: Beirowski
authors.forename: Simon A
authors.initials: SA
authors.lastname: Walker
authors.forename: Georg
authors.initials: G
authors.lastname: Haase
authors.forename: Arzu
authors.initials: A
authors.lastname: Celik
authors.forename: Robert
authors.initials: R
authors.lastname: Adalbert
authors.forename: Diana
authors.initials: D
authors.lastname: Wagner
authors.forename: Daniela
authors.initials: D
authors.lastname: Grumme
authors.forename: Richard R
authors.initials: RR
authors.lastname: Ribchester
authors.forename: Markus
authors.initials: M
authors.lastname: Plomann
authors.forename: Michael P
authors.initials: MP
authors.lastname: Coleman
mesh_terms: D000251:Adenosine Triphosphatases; D000595:Amino Acid Sequence; D000818:Animals; D019556:COS Cells; D018797:Cell Cycle Proteins / Q000737:chemistry / Q000378:metabolism*; D002467:Cell Nucleus / Q000378:metabolism*; D002478:Cells, Cultured; D002522:Chlorocebus aethiops; D019143:Evolution, Molecular; D006367:HeLa Cells; D006801:Humans; D042581:Intranuclear Space / Q000378:metabolism; D051379:Mice; D008810:Mice, Inbred C57BL; D008969:Molecular Sequence Data; D009419:Nerve Tissue Proteins / Q000737:chemistry / Q000378:metabolism*; D009612:Nicotinamide-Nucleotide Adenylyltransferase / Q000378:metabolism; D011485:Protein Binding; D021381:Protein Transport; D051381:Rats; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D025801:Ubiquitin / Q000378:metabolism; D000074405:Valosin Containing Protein
chemical_list: D018797:Cell Cycle Proteins; D009419:Nerve Tissue Proteins; D011993:Recombinant Fusion Proteins; D025801:Ubiquitin; C442580:Wld protein, mouse; D009612:Nicotinamide-Nucleotide Adenylyltransferase; C510972:Nmnat1 protein, mouse; D000251:Adenosine Triphosphatases; C000614178:VCP protein, human; D000074405:Valosin Containing Protein; C000614179:Vcp protein, mouse; C000614180:Vcp protein, rat