Mutations in the cytoplasmic domain of P0 reveal a role for PKC-mediated phosphorylation in adhesion and myelination.
Mutations in P0 (MPZ), the major myelin protein of the peripheral nervous system, cause the inherited demyelinating neuropathy Charcot-Marie-Tooth disease type 1B. P0 is a member of the immunoglobulin superfamily and functions as a homophilic adhesion molecule. We now show that point mutations in the cytoplasmic domain that modify a PKC target motif (RSTK) or an adjacent serine residue abolish P0 adhesion function and can cause peripheral neuropathy in humans. Consistent with these data, PKCalpha along with the PKC binding protein RACK1 are immunoprecipitated with wild-type P0, and inhibition of PKC activity abolishes P0-mediated adhesion. Point mutations in the RSTK target site that abolish adhesion do not alter the association of PKC with P0; however, deletion of a 14 amino acid region, which includes the RSTK motif, does abolish the association. Thus, the interaction of PKCalpha with the cytoplasmic domain of P0 is independent of specific target residues but is dependent on a nearby sequence. We conclude that PKC-mediated phosphorylation of specific residues within the cytoplasmic domain of P0 is necessary for P0-mediated adhesion, and alteration of this process can cause demyelinating neuropathy in humans.
Xu
W
W
Department of Neurology and the Center for Molecular Medicine and Genetics, Wayne State University Detroit, MI 48202, USA.
Shy
M
M
Kamholz
J
J
Elferink
L
L
Xu
G
G
Lilien
J
J
Balsamo
J
J
D000595:Amino Acid Sequence; D000596:Amino Acids; D000818:Animals; D001665:Binding Sites; D002448:Cell Adhesion / Q000502:physiology; D002607:Charcot-Marie-Tooth Disease / Q000235:genetics / Q000378:metabolism*; D003593:Cytoplasm / Q000378:metabolism; D003711:Demyelinating Diseases; D006367:HeLa Cells; D006801:Humans; D007527:Isoenzymes / Q000378:metabolism; D007739:L Cells; D051379:Mice; D008969:Molecular Sequence Data; D018993:Myelin P0 Protein / Q000235:genetics / Q000378:metabolism* / Q000502:physiology; D010455:Peptides / Q000378:metabolism; D010766:Phosphorylation; D011493:Protein Kinase C / Q000378:metabolism*; D051571:Protein Kinase C-alpha; D000075370:Receptors for Activated C Kinase; D017384:Sequence Deletion
D000596:Amino Acids; D007527:Isoenzymes; D018993:Myelin P0 Protein; D010455:Peptides; D000075370:Receptors for Activated C Kinase; C070301:peptide I; C496628:PRKCA protein, human; C496630:Prkca protein, mouse; D011493:Protein Kinase C; D051571:Protein Kinase C-alpha