Cloning, structural characterization, and chromosomal localization of the human orthologue of Saccharomyces cerevisiae MSH5 gene
Cloning, structural characterization, and chromosomal localization of the human orthologue of Saccharomyces cerevisiae MSH5 gene
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DOI：
10.1006/geno.1998.5374
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发表时间：
1998-08-15
期刊：
影响因子：
4.4
通讯作者：
Doggett, NA
中科院分区：
文献类型：
--
作者：
Her, C;Doggett, NA
我们已经克隆和鉴定了酿酒酵母MutS同源物5（MSH 5）cDNA的人类直系同源物，以及编码MSH 5 cDNA的人类基因，作为了解这种新型人类蛋白质生物学功能所涉及的分子遗传机制的一步。鉴定的cDNA含有2505-bp的opera阅读框架（ORF），其编码834个氨基酸的多肽，预测分子量为92.9 kDa。由该cDNA编码的氨基酸序列包括存在于细菌至人类中的所有已知MutS同源物中保守的序列基序。氨基酸序列分析表明，该cDNA属于MutS家族，与S的同源性为30%。酿酒酵母MSR 5是在减数分裂期间促进交换中起关键作用的酵母基因。北方印迹分析表明，在所有测试的人体组织中存在2.9 kb的人MSH 5 mRNA种类，但在人睾丸中表达最高，睾丸是一个含有进行恒定DNA合成和减数分裂的细胞的器官。人类MSH 5的表达模式与先前鉴定的人类MutS同源物MSH 2、MSH 3和MSH 6基因的表达模式相似，这些基因参与遗传性非息肉病性结直肠癌（HNPCC）的发病机制。为了加快寻找与这种新的人类MutS同源物相关的潜在疾病，我们还确定了人类MSH 5 Focus的染色体位置和结构。序列和结构表征表明，MSH 5跨度约25 kb，包含26个外显子，其长度范围从外显子8的36 bp到外显子25的254 bp。用荧光原位杂交技术将MSH 5定位于人染色体6p21.3带。对MSH 5的序列和基因结构的了解将使我们能够研究MSH 5在减数分裂和/或DNA复制错配修复中可能发挥的作用。(C)北京：科学出版社.
We have cloned and characterized the human orthologue of the Saccharomyces cerevisiae MutS homologue 5 (MSH5) cDNA, as well as the human gene that encodes the MSH5 cDNA, as a step toward understanding the molecular genetic mechanisms involved in the biological function of this novel human protein. The identified cDNA contains a 2505-bp opera reading frame (ORF) that encodes an 834-amino-acid polypeptide with a predicted molecular mass of 92.9 kDa. The amino acid sequence encoded by this cDNA includes sequence motifs that are conserved in all known MutS homologues existing in bacteria to humans. The cDNA appears, on the basis of amino acid sequence analysis, to be a member of the MutS family and shares 30% sequence identity with that of S. cerevisiae MSR5, a yeast gene that plays a critical role in facilitating crossover during meiosis. Northern blot analysis demonstrated the presence of a 2.9-kb human MSH5 mRNA species in all human tissues tested, but the highest expression was in human testis, an organ containing cells that undergo constant DNA synthesis and meiosis. The expression pattern of human MSH5 resembled that of the previously identified human MutS homologues MSH2, MSH3, and MSH6-genes that are involved in the pathogenesis of hereditary nonpolyposis colorectal cancer (HNPCC). In an effort to expedite the search for potential disease association with this new human MutS homologue, we have also determined the chromosomal location and structure of the human MSH5 Focus. Sequence and structural characterization demonstrated that MSH5 spans approximately 25 kb and contains 26 exons that range in length from 36 bp for exon 8 to 254 bp for exon 25. MSH5 has been mapped to human chromosome band 6p21.3 by fluorescence in situ hybridization. Knowledge of the sequence and gene structure of MSH5 will now enable studies of the possible roles MSH5 may play in meiosis and/or DNA replicative mismatch repair. (C) 1998 Academic Press.