Functional analysis of novel mutations in PAX9 associated with familial oligodontia.
- Author:
Ji-lin ZHAO
1
;
Yang-xi CHEN
;
Lang BAO
;
Tuo-jiang WU
;
Li ZHOU
Author Information
- Publication Type:Journal Article
- MeSH: Anodontia; genetics; Base Sequence; Chromatography, High Pressure Liquid; DNA Mutational Analysis; Electrophoretic Mobility Shift Assay; Family Health; Humans; Mutation; PAX9 Transcription Factor; genetics; metabolism; Polymerase Chain Reaction
- From: Chinese Journal of Medical Genetics 2005;22(4):419-422
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVETo gain new insights into the molecular pathogenesis of the 109(InsG) and 139(C--> T) mutations and their roles in familial oligodontia.
METHODSThe 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.
RESULTSWild 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.
CONCLUSIONThe 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.