Functional analysis of the human MCL-1 gene
Functional analysis of the human MCL-1 gene
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DOI：
10.1007/pl00000728
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发表时间：
2000-04-01
影响因子：
8
通讯作者：
Edwards, SW
中科院分区：
文献类型：
--
作者：
Akgul, C;Turner, PC;Edwards, SW
我们已经分离到一个6.5kb的人类基因组片段，该片段编码MCL-1基因。将该基因的编码区与已发表的全长基因进行比较，发现该基因含有3个外显子和2个内含子，3‘端非翻译区有370个碱基对。我们已经将一个主要的转录起始点定位到翻译起始密码子上游的80个残基。报告基因分析表明，佛波酯(PMA)刺激活性和粒细胞-巨噬细胞集落刺激因子(GM-CSF)刺激活性的调控序列位于转录起始点上游5‘侧翼区的前294个碱基内。产生了一个缺失突变体，在-215和-168残基之间缺失47bp：在该突变体中，7个GGCCCC重复序列中的6个和2个GCTCA重复序列被缺失。在该缺失突变体中，血清刺激和GM-CSF刺激的报告活性显著降低，PMA刺激的活性略有下降。虽然人类和小鼠基因的编码区和3‘非翻译区具有显著的序列相似性，但这两个物种的基因5’侧翼区的序列相似性很小。然而，在这两个基因中检测到了一些共同的转录因子结合位点的序列，表明在这些不同的物种中，转录可能受到相似的信号通路的调控。
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.