ZNF397, a new class of interphase to early prophase-specific, SCAN-zinc-finger, mammalian centromere protein
ZNF397, a new class of interphase to early prophase-specific, SCAN-zinc-finger, mammalian centromere protein
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DOI：
10.1007/s00412-008-0155-7
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发表时间：
2008-08-01
期刊：
影响因子：
1.6
通讯作者：
Kalitsis, P.
中科院分区：
文献类型：
--
作者：
Bailey, S. L.;Chang, S. C.;Kalitsis, P.
着丝粒是一种复杂的结构，需要在有丝分裂时使新合成的姐妹染色单体均匀分离。着丝粒研究的一个重要目标是确定构成着丝粒的完整蛋白质组分。在这里，我们使用一名西瓜胃病患者的着丝粒阳性自身免疫血清鉴定了一种新的着丝粒蛋白。Western印迹和噬菌体表达文库的筛选显示了一个60 kDa的蛋白ZNF397。该蛋白属于锌指蛋白超家族中经典的Cys(2)His(2)基团，包含两个保守结构域：一个富含亮氨酸的SCAN结构域和9个Cys(2)His(2)锌指。生物信息学分析表明，ZNF397在胎盘哺乳动物中是保守的。稳定的GFP：表达ZNF397的人类细胞在间期和早期阶段显示ZNF397与结构性着丝粒蛋白CENP-A共定位。缺失和结构域交换结构表明，扫描域对于着丝粒定位是必要的，但不是充分的。利用小鼠同源基因(Zfp397)对小鼠进行的基因敲除研究表明，ZNF397是一种非必需蛋白质。这些特性将ZNF397定义为一类新的间期到早期前期特异的包含SCAN结构域的哺乳动物着丝粒蛋白的成员。对该蛋白在着丝粒转录中的可能作用进行了讨论。
The centromere is a complex structure required for equal segregation of newly synthesised sister chromatids at mitosis. One of the significant objectives in centromere research is to determine the complete repertoire of protein components that constitute the kinetochore. Here, we identify a novel centromere protein using a centromere-positive autoimmune serum from a patient with watermelon stomach disease. Western blot and screening of a lambda phage expression library revealed a 60-kDa protein, ZNF397. This protein belongs to the classical Cys(2)His(2) group of the zinc-finger protein superfamily and contains two conserved domains: a leucine-rich SCAN domain and nine Cys(2)His(2) zinc fingers. Bioinformatic analysis shows that ZNF397 is conserved in placental mammals. Stable GFP:ZNF397-expressing human cells show co-localisation of ZNF397 with the constitutive centromere protein CENP-A during interphase and early prophase. Deletion and domain-swap constructs indicate that the SCAN domain is necessary but not sufficient for centromere localisation. Gene-knockout studies in mice using the mouse orthologue (Zfp397) reveal that ZNF397 is a non-essential protein. These properties define ZNF397 as a member of a new class of interphase to early prophase-specific and SCAN domain-containing mammalian centromere protein. The possible role of this protein in transcription at the centromere is discussed.