Down-regulation of cyclin B1 gene transcription in terminally differentiated skeletal muscle cells is associated with loss of functional CCAAT-binding NF-Y complex

A. Farina, I. Manni, G. Fontemaggi, M. Tiainen, C. Cenciarelli, M. Bellorini, R. Mantovani, A. Sacchi, G. Piaggio

Research output: Contribution to journalArticle

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Abstract

The observation that cyclin B1 protein and mRNAs are down-regulated in terminally differentiated (TD) C2C12 cells, suggested us to investigate the transcriptional regulation of the cyclin B1 gene in these cells. Transfections of cyclin B1 promoter constructs indicate that two CCAAT boxes support cyclin B1 promoter activity in proliferating cells. EMSAs demonstrate that both CCAAT boxes are recognized by the trimeric NF-Y complex in proliferating but not in TD cells. Transfecting a dominant-negative mutant of NF-YA we provide evidence that NF-Y is required for maximal promoter activity. Addition of recombinant NF-YA to TD C2C12 nuclear extracts restores binding activity in vitro, thus indicating that the loss of NF-YA in TD cells is responsible for the lack of the NF-Y binding to the CCAAT boxes. Consistent with this, we found that the NF-YA protein is absent in TD C2C12 cells. In conclusion, our data demonstrate that NF-Y is required for cyclin B1 promoter activity. We also demonstrate that cyclin B1 expression is regulated at the transcriptional level in TD C2C12 cells and that the switch-off of cyclin B1 promoter activity in differentiated cells depends upon the loss of a functional NF-Y complex. In particular the loss of NF-YA protein is most likely responsible for its inactivation.

Original languageEnglish
Pages (from-to)2818-2827
Number of pages10
JournalOncogene
Volume18
Issue number18
DOIs
Publication statusPublished - May 6 1999

Fingerprint

Cyclin B1
Muscle Cells
Skeletal Muscle
Down-Regulation
Genes
Proteins
Transfection
Messenger RNA

Keywords

  • CCAAT-binding factor NF-Y
  • Cyclin B1 promoter
  • Muscle differentiation

ASJC Scopus subject areas

  • Molecular Biology
  • Cancer Research
  • Genetics

Cite this

Down-regulation of cyclin B1 gene transcription in terminally differentiated skeletal muscle cells is associated with loss of functional CCAAT-binding NF-Y complex. / Farina, A.; Manni, I.; Fontemaggi, G.; Tiainen, M.; Cenciarelli, C.; Bellorini, M.; Mantovani, R.; Sacchi, A.; Piaggio, G.

In: Oncogene, Vol. 18, No. 18, 06.05.1999, p. 2818-2827.

Research output: Contribution to journalArticle

Farina, A. ; Manni, I. ; Fontemaggi, G. ; Tiainen, M. ; Cenciarelli, C. ; Bellorini, M. ; Mantovani, R. ; Sacchi, A. ; Piaggio, G. / Down-regulation of cyclin B1 gene transcription in terminally differentiated skeletal muscle cells is associated with loss of functional CCAAT-binding NF-Y complex. In: Oncogene. 1999 ; Vol. 18, No. 18. pp. 2818-2827.
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AU - Cenciarelli, C.

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