Essentiality of CENP-A Depends on Its Binding Mode to HJURP
Essentiality of CENP-A Depends on Its Binding Mode to HJURP
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DOI：
10.1016/j.celrep.2020.108388
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发表时间：
2020-11-17
期刊：
影响因子：
8.8
通讯作者：
Fukagawa, Tatsuo
中科院分区：
文献类型：
--
作者：
Hori, Tetsuya;Cao, JingHui;Fukagawa, Tatsuo
CENP-A掺入对于着丝粒特化是关键的，并且由伴侣HJURP介导。CENP-A的CENP-A靶向结构域（CATD）特异性结合HJURP，并且这种结合是保守的。然而，CENP-A-HJURP的结合界面尚未被理解。在这里，我们确定了鸡CENP-A或HJURP的关键残留物。鸡CENP-A的α-螺旋中的A59 Q突变导致CENP-A错误掺入和随后的细胞死亡，而人CENP-A中的相应突变则不会。我们还发现HJURP的W53是CENP-A中A59的接触位点，在鸡细胞中也是必需的。我们的综合分析表明，HJURP的亲和力CATD鸡和人之间的不同。然而，将两个精氨酸残基引入鸡HJURP α A-螺旋抑制了CENP-A在表达CENP-A（A59 Q）的鸡细胞中的错误掺入。我们的数据解释了CENP-A-HJURP相互作用的CENP-A本质的机制和演变。
CENP-A incorporation is critical for centromere specification and is mediated by the chaperone HJURP. The CENP-A-targeting domain (CATD) of CENP-A specifically binds to HJURP, and this binding is conserved. However, the binding interface of CENP-A-HJURP is yet to be understood. Here, we identify the critical residues for chicken CENP-A or HJURP. The A59Q mutation in the alpha-helix of chicken CENP-A causes CENP-A mis-incorporation and subsequent cell death, whereas the corresponding mutation in human CENP-A does not. We also find that W53 of HJURP, which is a contact site of A59 in CENP-A, is also essential in chicken cells. Our comprehensive analyses reveal that the affinities of HJURP to CATD differ between chickens and humans. However, the introduction of two arginine residues to the chicken HJURP alpha A-helix suppresses CENP-A mis-incorporation in chicken cells expressing CENP-A(A59Q). Our data explain the mechanisms and evolution of CENP-A essentiality by the CENP-A-HJURP interaction.