DNA binding of centromere protein C (CENPC) is stabilized by single-stranded RNA.
DNA binding of centromere protein C (CENPC) is stabilized by single-stranded RNA.
复制标题
DOI：
10.1371/journal.pgen.1000835
复制
发表时间：
2010-02-05
期刊：
影响因子：
4.5
通讯作者：
Dawe RK
中科院分区：
文献类型：
--
作者：
Du Y;Topp CN;Dawe RK
着丝粒是基因组和细胞骨架之间的附着点：着丝粒与动粒结合，动粒又与纺锤体结合并移动染色体。巧合的是，着丝粒的DNA序列在使动粒永久化方面几乎没有作用。因此，它们是遗传信息以独立于DNA序列的方式（表观遗传学）传播的突出例子。已经发现，从着丝粒转录的RNA保持结合在动粒区域内，并且这种RNA的局部群体被认为是表观遗传标记系统的一部分。在这里，我们进行了玉米CENPC，一个关键的内部动粒蛋白的遗传和生化研究。我们表明，DNA结合是由一个122个氨基酸长的局部区域赋予的，并且DNA结合反应对单链RNA非常敏感。长的单链核酸强烈促进CENPC与DNA的结合，并且稳定DNA结合的RNA类型在大小和特征上与体内着丝粒上存在的RNA相匹配。用HIV整合酶结合结构域去除或替换结合模块导致CENPC在体内部分离域。这些数据表明，着丝粒RNA通过改变其DNA结合特性有助于将CENPC招募到内部动粒。在这里，我们解决的问题，遗传信息是如何从一代传递到下一代没有特定的DNA序列的参与。这种类型的遗传被称为表观遗传学。着丝粒序列是高度可变的，并且在许多情况下不足以用于着丝粒功能。相反，DNA的次要特征，如甲基化或相关的RNA分子可能有助于招募关键的着丝粒结合蛋白。来自几个物种的先前数据已经确定，单链RNA在着丝粒染色质上惊人地丰富。在这里，我们确定了玉米（CENPC）中的一个关键着丝粒结合蛋白的DNA结合域，并表明它需要单链RNA才能在体外有效地结合DNA。当DNA/RNA结合结构域被删除时，CENPC靶向着丝粒的准确性降低，但没有消除。这些结果支持了这样一种观点，即着丝粒结合的RNA是确保着丝粒稳定遗传的表观遗传决定过程的一个组成部分。此外，我们的数据表明了RNA如何影响染色质蛋白与DNA结合的一般机制。
Centromeres are the attachment points between the genome and the cytoskeleton: centromeres bind to kinetochores, which in turn bind to spindles and move chromosomes. Paradoxically, the DNA sequence of centromeres has little or no role in perpetuating kinetochores. As such they are striking examples of genetic information being transmitted in a manner that is independent of DNA sequence (epigenetically). It has been found that RNA transcribed from centromeres remains bound within the kinetochore region, and this local population of RNA is thought to be part of the epigenetic marking system. Here we carried out a genetic and biochemical study of maize CENPC, a key inner kinetochore protein. We show that DNA binding is conferred by a localized region 122 amino acids long, and that the DNA-binding reaction is exquisitely sensitive to single-stranded RNA. Long, single-stranded nucleic acids strongly promote the binding of CENPC to DNA, and the types of RNAs that stabilize DNA binding match in size and character the RNAs present on kinetochores in vivo. Removal or replacement of the binding module with HIV integrase binding domain causes a partial delocalization of CENPC in vivo. The data suggest that centromeric RNA helps to recruit CENPC to the inner kinetochore by altering its DNA binding characteristics. Here we address the issue of how genetic information is passed from one generation to the next without the involvement of specific DNA sequences. This type of inheritance is referred to as epigenetics. Centromeric sequences are highly variable and in many cases are not sufficient for centromere function. Rather, secondary features of the DNA, such as methylation or associated RNA molecules may serve to recruit key centromere binding proteins. Prior data from several species have established that single-stranded RNAs are surprisingly abundant on centromeric chromatin. Here we identified the DNA-binding domain of a key centromere binding protein in maize (CENPC) and showed that it requires single-stranded RNA to effectively bind DNA in vitro. When the DNA/RNA binding domain was deleted, the accuracy of CENPC targeting to centromeres was reduced but not abolished. The results bolster the view that centromere-bound RNA is one component of the epigenetic determination process that assures centromeres are stably inherited. In addition, our data suggest a general mechanism for how RNA can influence the binding of chromatin proteins to DNA.
登录
查看更多内容
影响因子：
4.5
作者：
Chueh, Anderly C.;Northrop, Emma L.;Brettingham-Moore, Kate H.;Choo, K. H. Andy;Wong, Lee H.
通讯作者：
Wong, Lee H.
影响因子：
4.5
作者：
Blower MD;Daigle T;Kaufman T;Karpen GH
通讯作者：
Karpen GH
影响因子：
11.6
作者：
Dawe, RK;Reed, LM;Hiatt, EN
通讯作者：
Hiatt, EN
影响因子：
64.5
作者：
Hori, Tetsuya;Amano, Miho;Fukagawa, Tatsuo
通讯作者：
Fukagawa, Tatsuo
DOI：
10.1073/pnas.97.3.1148
发表时间：
2000-02-01
影响因子：
11.1
作者：
Howman, EV;Fowler, KJ;Choo, KHA
通讯作者：
Choo, KHA