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H.-G. Chen, J. Qin, Q. Yan, J.-P. Liu, W.-B. Liu, Y. Liu, D. W.-C. Li; Dissection of the Promoter Region of the Gene Encoding the Regulatory A-alpha Subunit of the Protein Phosphatase-2a. Invest. Ophthalmol. Vis. Sci. 2007;48(13):4239.
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Protein serine/threonine phosphatases play important role in regulation gene expression, cell proliferation, differentiation, and apoptosis. The protein serine/threonine phosphatase-2A (PP-2A) is one of the major serine/threonine phosphatases in the ocular tissues. The function of PP-2A is largely regulated by the two families of regulatory subunits: A and B. In the present study, we have cloned the promoter region of the A regulatory subunit (alpha form) and dissected its regulatory regions.
Bioinformatics and PCR were used for the cloning of the promoter region (1954 bp-named A1) of PP-2A-Aalpha. This promoter region was sequentially deleted into 4 fragments named A2 (1526), A3 (1106), A4 (773 bp) and A5 (677 bp). In vitro mutagenesis was used to conduct the point mutation to identify the cis-elements in the A5 region. Different promoter fragments and point mutation mutants were linked to luciferase reporter gene for their activity analysis in rabbit lens epithelial cells.
Reporter gene activity assays reveal that there exists a repressor between 1954 bp and 1106 bp. Within the first 677 bp (A5) of the promoter region, SP-1 plays a vital role to maintain expression from the PP-2A-Aalpha promoter. In contrast, AP-2 seems to act as a repressor in this region (A5).
Multiple cis-regulatory elements are present in the promoter region of PP-2A-Aalpha. Among these elements, SP-1 and AP-2 play important roles.
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