Repression of transcription of the p27(Kip1) cyclin-dependent kinase inhibitor gene by c-Myc.
Upon engagement of the B Cell Receptor (BCR) of WEHI 231 immature B cells, a drop in c-Myc expression is followed by activation of the cyclin-dependent kinase inhibitor (CKI) p27(Kip1), which induces growth arrest and apoptosis. Here, we report inverse patterns of p27 and c-Myc protein expression follow BCR engagement. We present evidence demonstrating, for the first time, that the p27(Kip1) gene is a target of transcriptional repression by c-Myc. Specifically, the changes in p27 protein levels correlated with changes in p27 mRNA levels, and gene transcription. Induction of p27 promoter activity followed BCR engagement of WEHI 231 cells, and this induction could be repressed upon co-transfection of a c-Myc expression vector. Inhibition of the TATA-less p27 promoter by c-Myc was also observed in Jurkat T cells, vascular smooth muscle, and Hs578T breast cancer cells, extending the observation beyond immune cells. Consistent with a putative Inr element CCAGACC (where +1 is underlined) at the start site of transcription in the p27 promoter, deletion of Myc homology box II reduced the extent of repression. Furthermore, enhanced repression was observed upon transfection of the c-Myc 'super-repressor', with mutation of Phe115 to Leu. The sequences mediating transcriptional activity and c-Myc repression were mapped to bp -20 to +20 of the p27 gene. Finally, binding of Max was shown to facilitate c-Myc binding and repression of p27 promoter activity. Overall, these studies identify the p27 CKI gene as a new target whereby c-Myc can control cell proliferation, survival and neoplastic transformation.
Yang
W
W
Department of Biochemistry, Boston University Medical School, Boston, Maryland, MA 02118, USA.
Shen
J
J
Wu
M
M
Arsura
M
M
FitzGerald
M
M
Suldan
Z
Z
Kim
D W
DW
Hofmann
C S
CS
Pianetti
S
S
Romieu-Mourez
R
R
Freedman
L P
LP
Sonenshein
G E
GE
D000818:Animals; D000888:Antibodies, Anti-Idiotypic / Q000276:immunology; D051778:Basic Helix-Loop-Helix Leucine Zipper Transcription Factors; D050976:Basic-Leucine Zipper Transcription Factors; D018797:Cell Cycle Proteins; D002471:Cell Transformation, Neoplastic; D002478:Cells, Cultured; D050760:Cyclin-Dependent Kinase Inhibitor p27; D004268:DNA-Binding Proteins / Q000378:metabolism; D016193:G1 Phase; D006801:Humans; D008869:Microtubule-Associated Proteins / Q000235:genetics*; D009131:Muscle, Smooth, Vascular / Q000378:metabolism; D011401:Promoter Regions, Genetic; D016271:Proto-Oncogene Proteins c-myc / Q000502:physiology*; D012333:RNA, Messenger / Q000032:analysis; D011947:Receptors, Antigen, B-Cell / Q000502:physiology; D012097:Repressor Proteins / Q000502:physiology*; D014157:Transcription Factors; D025521:Tumor Suppressor Proteins
D000888:Antibodies, Anti-Idiotypic; D051778:Basic Helix-Loop-Helix Leucine Zipper Transcription Factors; D050976:Basic-Leucine Zipper Transcription Factors; D018797:Cell Cycle Proteins; D004268:DNA-Binding Proteins; C499330:MAX protein, human; D008869:Microtubule-Associated Proteins; C067850:Myc associated factor X; D016271:Proto-Oncogene Proteins c-myc; D012333:RNA, Messenger; D011947:Receptors, Antigen, B-Cell; D012097:Repressor Proteins; D014157:Transcription Factors; D025521:Tumor Suppressor Proteins; C044203:anti-IgM; D050760:Cyclin-Dependent Kinase Inhibitor p27