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首页> 外文期刊>Molecular and Cellular Biology >CENP-B binds a novel centromeric sequence in the Asian mouse Mus caroli.
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CENP-B binds a novel centromeric sequence in the Asian mouse Mus caroli.

机译：CENP-B结合亚洲小鼠Mus caroli中的新型着丝粒序列。

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Minor satellite DNA, found at Mus musculus centromeres, is not present in the genome of the Asian mouse Mus caroli. This repetitive sequence family is speculated to have a role in centromere function by providing an array of binding sites for the centromere-associated protein CENP-B. The apparent absence of CENP-B binding sites in the M. caroli genome poses a major challenge to this hypothesis. Here we describe two abundant satellite DNA sequences present at M. caroli centromeres. These satellites are organized as tandem repeat arrays, over 1 Mb in size, of either 60- or 79-bp monomers. All autosomes carry both satellites and small amounts of a sequence related to the M. musculus major satellite. The Y chromosome contains small amounts of both major satellite and the 60-bp satellite, whereas the X chromosome carries only major satellite sequences. M. caroli chromosomes segregate in M. caroli x M. musculus interspecific hybrid cell lines, indicating that the two sets of chromosomes can interact with the same mitotic spindle. Using a polyclonal CENP-B antiserum, we demonstrate that M. caroli centromeres can bind murine CENP-B in such an interspecific cell line, despite the absence of canonical 17-bp CENP-B binding sites in the M. caroli genome. Sequence analysis of the 79-bp M. caroli satellite reveals a 17-bp motif that contains all nine bases previously shown to be necessary for in vitro binding of CENP-B. This M. caroli motif binds CENP-B from HeLa cell nuclear extract in vitro, as indicated by gel mobility shift analysis. We therefore suggest that this motif also causes CENP-B to associate with M. caroli centromeres in vivo. Despite the sequence differences, M. caroli presents a third, novel mammalian centromeric sequence producing an array of binding sites for CENP-B.
机译：在小家鼠着丝粒中发现的次要卫星DNA，在亚洲小鼠家鼠的基因组中不存在。通过提供与着丝粒相关蛋白CENP-B的一系列结合位点，推测该重复序列家族在着丝粒功能中起作用。在卡罗氏支原体基因组中显然没有CENP-B结合位点对该假设提出了重大挑战。在这里，我们描述了存在于卡罗莫氏丝状体的两个丰富的卫星DNA序列。这些卫星组织成60或79 bp单体的串联重复阵列，大小超过1 Mb。所有常染色体都携带小卫星和与小家鼠主要卫星有关的少量序列。 Y染色体包含少量主要卫星和60 bp卫星，而X染色体仅携带主要卫星序列。卡罗氏支原体染色体在卡罗氏支原体x小家鼠种间杂种细胞系中分离，表明两组染色体可以与同一有丝分裂纺锤体相互作用。使用多克隆CENP-B抗血清，我们证明尽管在M. caroli基因组中不存在规范的17 bp CENP-B结合位点，但Cararo着丝粒体可以在这种种间细胞系中结合鼠CENP-B。对该79 bp的卡罗氏疟原虫卫星进行的序列分析揭示了一个17 bp的基序，其中包含先前显示的9个碱基对体外结合CENP-B所必需的9个碱基。如凝胶迁移率移动分析所表明的，该卡罗氏疟原虫基序在体外结合来自HeLa细胞核提取物的CENP-B。因此，我们建议该基序还导致CENP-B在体内与卡罗氏支原体着丝粒相关。尽管序列不同，但卡罗氏支原体仍呈现出第三种新颖的哺乳动物着丝粒序列，产生了CENP-B结合位点阵列。

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