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Functional analysis of the human MCL-1 gene

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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.

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Received 20 January 2000; received after revision 28 February 2000; accepted 28 February 2000

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Akgul, C., Turner, P., White, M. et al. Functional analysis of the human MCL-1 gene . CMLS, Cell. Mol. Life Sci. 57, 684–691 (2000).

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