Cellular and Molecular Life Sciences CMLS

, Volume 57, Issue 4, pp 684–691 | Cite as

Functional analysis of the human MCL-1 gene

  • C. Akgul
  • P. C. Turner
  • M. R. H. White
  • S. W. Edwards*

Abstract.

We have isolated a 6.5-kb human genomic fragment that encodes the MCL-1 gene. Comparison of the coding region with the published full-length cDNA reveals that the gene contains three exons and two introns, and that our clone contains 370 bp of the 3′-untranslated region. We have mapped a major transcriptional start site to 80 residues upstream of the translation initiation codon. Reporter gene assays indicate that regulatory sequences responsible for phorbol ester (PMA)-stimulated activity and granulocyte-macrophage-colony-stimulating factor (GM-CSF)-stimulated activity were located within the first 294 bp of the 5′-flanking region upstream from the transcription start site. A deletion mutant was generated that lacked 47 bp between residues −215 and −168: in this mutant, six out of seven GGCCCC repeats and two GCTCA repeats were deleted. Serum-stimulated and GM-CSF-stimulated reporter activity were greatly decreased in this deletion mutant and PMA-stimulated activity was slightly decreased. While the coding and 3′-untranslated regions of the human and mouse genes have significant sequence similarity, there was very little sequence similarity in the 5′-flanking regions of the genes from these two species. Nevertheless, some consensus sequences for a number of transcription-factor-binding sites were detected in the two genes, indicating that transcription may be regulated by similar signalling pathways in these different species.

Key words. Apoptosis; Bcl-2 family; neutrophil; transcription; U-937; promoter; luciferase. 

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Copyright information

© Birkhäuser Verlag Basel, 2000

Authors and Affiliations

  • C. Akgul
    • 1
  • P. C. Turner
    • 1
  • M. R. H. White
    • 1
  • S. W. Edwards*
    • 1
  1. 1.School of Biological Sciences, Life Sciences Building, University of Liverpool, Liverpool L69 7ZB (UK), Fax +44 151 794 4349, e-mail: sbir12@liverpool.ac.ukUK

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