[Functional analysis of novel mutations in PAX9 associated with familial oligodontia].
OBJECTIVE
To gain new insights into the molecular pathogenesis of the 109(InsG) and 139(C--> T) mutations and their roles in familial oligodontia.


METHODS
The region of PAX9 paired domain (PAX9PD) was amplified and the expression plasmids were constructed in pGEXlambda -1T by PCR-based cloning. PAX9PD proteins were prepared on the basis of GST instruction. The binding of wild type and two novely mutant PAX9 paired domain to double-stranded DNA targets were analyzed by gel mobility shift assay.


RESULTS
Wild type PAX9PD protein bind to the high affinity paired domain recognition sequences, CD19-2(A-ins) and Pax6CON, the 109(InsG) and 139(C--> T) mutant PAX9PD protein were unable to bind to these cognate DNA-binding sites.


CONCLUSIONS
The functional defects in DNA binding of mutant 109(InsG) PAX9 and 139(C--> T) PAX9, as well as loss-of-function of PAX9 most likely result in its haploinsufficiency during the patterning of dentition and the subsequent loss of posterior teeth.
Zhao
Ji-lin
JL
Department of Orthodontics, West China College of Stomatology, Sichuan University, Chengdu, Sichuan, 610041 PR China.
Chen
Yang-xi
YX
Bao
Lang
L
Wu
Tuo-jiang
TJ
Zhou
Li
L
D000848:Anodontia / Q000235:genetics*; D001483:Base Sequence; D002851:Chromatography, High Pressure Liquid; D004252:DNA Mutational Analysis; D024202:Electrophoretic Mobility Shift Assay; D005192:Family Health; D006801:Humans; D009154:Mutation; D051764:PAX9 Transcription Factor / Q000235:genetics* / Q000378:metabolism; D016133:Polymerase Chain Reaction
D051764:PAX9 Transcription Factor