title: Prenylation-defective human connexin32 mutants are normally localized and function equivalently to wild-type connexin32 in myelinating Schwann cells.
abstract: Mutations in GJB1, the gene encoding the gap junction protein connexin32 (Cx32), cause the X-linked form of Charcot-Marie-Tooth disease, an inherited demyelinating neuropathy. The C terminus of human Cx32 contains a putative prenylation motif that is conserved in Cx32 orthologs. Using [3H]mevalonolactone ([3H]MVA) incorporation, we demonstrated that wild-type human connexin32 can be prenylated in COS7 cells, in contrast to disease-associated mutations that are predicted to disrupt the prenylation motif. We generated transgenic mice that express these mutants in myelinating Schwann cells. Male mice expressing a transgene were crossed with female Gjb1-null mice; the male offspring were all Gjb1-null, and one-half were transgene positive; in these mice, all Cx32 was derived from expression of the transgene. The mutant human protein was properly localized in myelinating Schwann cells in multiple transgenic lines and did not alter the localization of other components of paranodes and incisures. Finally, both the C280G and the S281x mutants appeared to "rescue" the phenotype of Gjb1-null mice, because transgene-positive male mice had significantly fewer abnormally myelinated axons than did their transgene-negative male littermates. These results indicate that Cx32 is prenylated, but that prenylation is not required for proper trafficking of Cx32 and perhaps not even for certain aspects of its function, in myelinating Schwann cells.
authors.affiliation: Department of Neurology, The University of Pennsylvania Medical Center, Philadelphia, Pennsylvania 19104, USA. yanhuang@mail.med.upenn.edu
authors.forename: Yan
authors.initials: Y
authors.lastname: Huang
authors.forename: Erich E
authors.initials: EE
authors.lastname: Sirkowski
authors.forename: John T
authors.initials: JT
authors.lastname: Stickney
authors.forename: Steven S
authors.initials: SS
authors.lastname: Scherer
mesh_terms: D020816:Amino Acid Motifs / Q000235:genetics; D000818:Animals; D018388:Codon, Terminator; D017630:Connexins / Q000172:deficiency / Q000235:genetics* / Q000378:metabolism / Q000502:physiology*; D017124:Conserved Sequence; D003545:Cysteine; D004247:DNA / Q000378:metabolism; D019143:Evolution, Molecular; D005260:Female; D005267:Femoral Nerve / Q000502:physiology; D005998:Glycine; D006801:Humans; D008297:Male; D051379:Mice; D008822:Mice, Transgenic; D009154:Mutation; D009186:Myelin Sheath / Q000502:physiology*; D010641:Phenotype; D017368:Protein Prenylation / Q000235:genetics*; D012583:Schwann Cells / Q000502:physiology*; D012584:Sciatic Nerve / Q000502:physiology; D012694:Serine; D014018:Tissue Distribution; D000097002:Gap Junction beta-1 Protein
chemical_list: D018388:Codon, Terminator; D017630:Connexins; D003545:Cysteine; D004247:DNA; D005998:Glycine; D012694:Serine; D000097002:Gap Junction beta-1 Protein