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An unusual class of PITX2 mutations in axenfeld-rieger syndrome
Journal article   Peer reviewed

An unusual class of PITX2 mutations in axenfeld-rieger syndrome

Irfan SAADI, Rafael TORO, Adisa KUBURAS, Elena SEMINA, Jeffrey C MURRAY and Andrew F RUSSO
Birth defects research. A Clinical and molecular teratology, Vol.76(3), pp.175-181
2006
DOI: 10.1002/bdra.20226
PMCID: PMC4023635
PMID: 16498627

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Abstract

Background: Mutations in the PITX2 homeobox gene are known to contribute to Axenfeld-Rieger syndrome (ARS), an autosomal-dominant developmental disorder. Although most mutations are in the homeodomain and result in a loss of function, there is a growing subset in the C-terminal domain that has not yet been characterized. These mutations are of particular interest because the C-terminus has both inhibitory and stimulatory activities. Methods: In this study we used a combination of in vitro DNA binding and transfection reporter assays to investigate the fundamental issue of whether C-terminal mutations result in gain or loss of function at a cellular level. Results: We report a new frameshift mutation in the PITX2 allele that predicts a truncated protein lacking most of the C-terminal domain (D122FS). This newly reported mutant and another ARS C-terminal mutant (W133Stop) both have greater binding than wild-type to the bicoid element. Of interest, the mutants yielded approximately 5-fold greater activation of the prolactin promoter in CHO cells, even though the truncated proteins were expressed at lower levels than the wild-type protein. The truncated proteins also had greater than wild-type activity in 2 other cell lines, including the LS8 oral epithelial line that expresses the endogenous Pitx2 gene. Conclusions: The results indicate that the PITX2 C-terminal domain has inhibitory activity and support the notion that ARS may also be caused by gain-of-function mutations.
Ophthalmology Biological and medical sciences Medical sciences Malformations of the eye

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