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Nucleic Acids Research, 2000, Vol. 28, No. 22 4410-4418
© 2000 Oxford University Press

The tumour suppressor protein p53 can repress transcription of cyclin B

Karen Krause, Mark Wasner, Wibke Reinhard, Ulrike Haugwitz, Christine Lange-zu Dohna, Joachim Mössner and Kurt Engeland*

Medizinische Klinik II, Max Bürger Forschungszentrum, Universität Leipzig, Johannisallee 30, D-04103 Leipzig, Germany

The tumour suppressor protein p53 has functions in controlling the G1/S and G2/M transitions. Central regulators for progression from G2 to mitosis are B-type cyclins complexed with cdc2 kinase. In mammals two cyclin B proteins are found, cyclin B1 and B2. We show that upon treatment of HepG2 cells with 5-fluorouracil or methotrexate, p53 levels increase while concentrations of cyclin B2 mRNA, measured by RT–PCR with the LightCycler system, are reduced. In DLD-1 colorectal adenocarcinoma cells (DLD-1-tet-off-p53) cyclin B1 and B2 mRNA levels drop after expression of wild-type p53 but not after induction of a DNA binding-deficient mutant of p53. Analysis of the cyclin B2 promoter reveals specific repression of this gene by p53. Transfection of wild-type p53 into SaOS-2 cells shuts off transcription from a cyclin B2 promoter–luciferase construct whereas a p53 mutant protein does not. The cyclin B2 promoter does not contain a consensus p53 binding site. Most of the p53-dependent transcriptional responsiveness resides in its 226 bp core promoter. Taken together with earlier observations on p53-dependent transcription of cyclin B1, our results suggest that one way of regulating G2 arrest may be a reduction in cyclin B levels through p53-dependent transcriptional repression.

* To whom correspondence should be addressed. Tel: +49 341 972 5900; Fax: +49 341 971 2209; Email: engeland{at}medizin.uni-leipzig.de


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