title: The Boston-type craniosynostosis mutation MSX2 (P148H) results in enhanced susceptibility of MSX2 to ubiquitin-dependent degradation.
abstract: Boston-type craniosynostosis is caused by a single amino acid substitution, P148H, in the transcription factor MSX2. The increased binding affinity of MSX2 (P148H) to the response element has led many to hypothesize that the substitution is a gain-of-function mutation. However, there have been conflicting reports on the function of MSX2, and by extension, the nature of the P148H mutation remains unclear. In this study, we have examined the molecular mechanism of MSX2 function and the nature of the P148H mutation. During cranial suture closure of rodent, Msx2 expression was detected in the suture space. Overexpression of wild type MSX2 in mesenchymal cells stimulated cell proliferation and cyclin D1 expression, whereas P148H mutant did not. These results indicated that MSX2 is involved in maintaining the suture space by stimulating suture mesenchymal cell proliferation and that P148H is defective in this process. The protein levels of P148H were lower than wild type Msx2 (Msx2-WT), and pulse-chase experiments indicated that the mutant protein has a shorter half-life than the Msx2-WT protein. The ubiquitylation level of P148H was greater than that of Msx2-WT. The degradation of Msx2 was mediated by Praja1, and the P148H mutant was degraded more effectively than WT. The ubiquitylation of Msx2-WT was higher in the presence of Msx2 (P148H), which indicated that P148H functions as a dominant-negative mutant. Collectively, the primary function of MSX2 in suture closure is the induction of cell proliferation and suture maintenance, and the mutation results in an increased susceptibility of both wild type and mutant MSX2 to proteasomal degradation.
authors.affiliation: Department of Cell and Developmental Biology, School of Dentistry and Dental Research Institute, Seoul National University, Seoul, 110-749, Korea.
authors.forename: Won-Joon
authors.initials: WJ
authors.lastname: Yoon
authors.forename: Young-Dan
authors.initials: YD
authors.lastname: Cho
authors.forename: Kwang-Hwi
authors.initials: KH
authors.lastname: Cho
authors.forename: Kyung-Mi
authors.initials: KM
authors.lastname: Woo
authors.forename: Jeong-Hwa
authors.initials: JH
authors.lastname: Baek
authors.forename: Je-Yoel
authors.initials: JY
authors.lastname: Cho
authors.forename: Gwan-Shik
authors.initials: GS
authors.lastname: Kim
authors.forename: Hyun-Mo
authors.initials: HM
authors.lastname: Ryoo
mesh_terms: D002454:Cell Differentiation; D045744:Cell Line, Tumor; D049109:Cell Proliferation; D003398:Craniosynostoses / Q000235:genetics*; D019938:Cyclin D1 / Q000096:biosynthesis; D005786:Gene Expression Regulation; D020022:Genetic Predisposition to Disease; D018398:Homeodomain Proteins / Q000235:genetics* / Q000502:physiology; D006801:Humans; D017403:In Situ Hybridization; D008648:Mesoderm / Q000166:cytology; D008954:Models, Biological; D009154:Mutation; D010957:Plasmids / Q000378:metabolism; D025801:Ubiquitin / Q000378:metabolism*
chemical_list: D018398:Homeodomain Proteins; C084119:MSX2 protein; D025801:Ubiquitin; D019938:Cyclin D1