Mitotic phosphorylation regulates Hsp72 spindle localization by uncoupling ATP binding from substrate release.
Mitotic phosphorylation regulates Hsp72 spindle localization by uncoupling ATP binding from substrate release.
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DOI：
10.1126/scisignal.aao2464
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发表时间：
2018-08-14
影响因子：
7.3
通讯作者：
Bayliss R
中科院分区：
文献类型：
--
作者：
Mukherjee M;Sabir S;O'Regan L;Sampson J;Richards MW;Huguenin-Dezot N;Ault JR;Chin JW;Zhuravleva A;Fry AM;Bayliss R
Hsp 72是分子伴侣的70-kD热休克家族（Hsp 70）的成员，其包含通过连接子连接的核苷酸结合结构域（NBD）和底物结合结构域（SBD），所述连接子将二磷酸腺苷（ADP）交换为三磷酸腺苷（ATP）与蛋白质底物的释放偶联。在有丝分裂中，位于NBD中的Thr 66处的激酶NEK 6对Hsp 72的磷酸化促进Hsp 72定位于有丝分裂纺锤体，并且是有效的纺锤体组装和染色体聚集和分离所需的。在这里，我们确定了热休克蛋白72 NBD的晶体结构，含有遗传编码的磷酸丝氨酸在位置66。这揭示了稳定NBD内亚结构域之间相互作用的结构变化。ATP与未修饰的Hsp 72的NBD结合导致底物从SBD释放，但磷酸化的Hsp 72在ATP存在下保留底物。阻止磷酸化依赖性亚结构域相互作用的突变恢复了ATP结合和底物释放之间的联系。因此，Thr 66的磷酸化是一种可逆的机制，它使核苷酸结合和底物释放之间的变构连接发生变化，从而进一步了解Hsp 70家族的调控。我们建议，在有丝分裂过程中由NEK 6的Thr 66上的Hsp 72的磷酸化通过稳定其与有丝分裂纺锤体的组分的相互作用促进其在纺锤体上的定位。
Hsp72 is a member of the 70-kD heat shock family of molecular chaperones (Hsp70s) that comprise a nucleotide-binding domain (NBD) and a substrate-binding domain (SBD) connected by a linker that couples the exchange of adenosine diphosphate (ADP) for adenosine triphosphate (ATP) with the release of the protein substrate. Mitotic phosphorylation of Hsp72 by the kinase NEK6 at Thr66 located in the NBD promotes the localization of Hsp72 to the mitotic spindle and is required for efficient spindle assembly and chromosome congression and segregation. Here, we determined the crystal structure of the Hsp72 NBD containing a genetically encoded phosphoserine at position 66. This revealed structural changes that stabilized interactions between subdomains within the NBD. ATP binding to the NBD of unmodified Hsp72 resulted in the release of substrate from the SBD, but phosphorylated Hsp72 retained substrate in the presence of ATP. Mutations that prevented phosphorylation-dependent subdomain interactions restored the connection between ATP binding and substrate release. Thus, phosphorylation of Thr66 is a reversible mechanism that decouples the allosteric connection between nucleotide binding and substrate release, providing further insight into the regulation of the Hsp70 family. We propose that phosphorylation of Hsp72 on Thr66 by NEK6 during mitosis promotes its localization to the spindle by stabilizing its interactions with components of the mitotic spindle.
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影响因子：
7.3
作者：
Alexander J;Lim D;Joughin BA;Hegemann B;Hutchins JR;Ehrenberger T;Ivins F;Sessa F;Hudecz O;Nigg EA;Fry AM;Musacchio A;Stukenberg PT;Mechtler K;Peters JM;Smerdon SJ;Yaffe MB
通讯作者：
Yaffe MB
影响因子：
5.5
作者：
Fry AM;Bayliss R;Roig J
通讯作者：
Roig J
影响因子：
5.6
作者：
Bhattacharya, Akash;Kurochkin, Alexander V.;Yip, Grover N. B.;Zhang, Yongbo;Bertelsen, Eric B.;Zuiderweg, Erik R. P.
通讯作者：
Zuiderweg, Erik R. P.
影响因子：
16
作者：
Kityk, Roman;Kopp, Juergen;Mayer, Matthias P.
通讯作者：
Mayer, Matthias P.
DOI：
10.1107/s0907444904019158
发表时间：
2004-12-01
影响因子：
2.2
作者：
Emsley, P;Cowtan, K
通讯作者：
Cowtan, K