Wee1 protein kinase represses CDK1 required for G(2)/M transition. The mouse wee1 promoter contains multiple CACCC-boxes between -306 and +1 that can bind to Krüppel-like factor 3 (KLF3) transcriptional repressor. We found that increasing amounts of intracellular KLF3 decreased the amount of wee1 mRNA. A promoter reporter assay demonstrated that wee1 promoter activity was repressed by KLF3 overexpression. Elimination of the first and fourth CACCC-boxes suppressed KLF3-governed transcriptional repression. A gel-shift assay demonstrated that KLF3 binds to the first, third, and fourth CACCC-boxes with various strengths. Moreover, KLF3 was suggested to interact with the wee1 regulatory region in a physiological condition. Therefore, we concluded that KLF3 is a transcriptional repressor for wee1 gene. In a previous study, we demonstrated that TBP-like protein (TLP) inhibits wee1 promoter function. In this study, we found that the chromosomal wee1 gene is also down-regulated by KLF3. Since KLF3-repressed wee1 promoter function was further inhibited by TLP overexpression regardless of the inhibition degree of KLF3, we propose that TLP and KLF3 repress wee1 promoter independently.
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