Coordination of adjacent domains mediates TACC3-ch-TOG-clathrin assembly and mitotic spindle binding.

Coordination of adjacent domains mediates TACC3-ch-TOG-clathrin assembly and mitotic spindle binding.
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DOI:
10.1083/jcb.201211127
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发表时间:
2013-08-05
期刊:
The Journal of cell biology
影响因子:
--
通讯作者:
Royle SJ
Royle SJ
中科院分区:
其他
文献类型:
--
作者:
Hood FE;Williams SJ;Burgess SG;Richards MW;Roth D;Straube A;Pfuhl M;Bayliss R;Royle SJ

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磷酸化诱导的TACC3和网格蛋白相互作用协调每个蛋白的相邻结构域,形成微管结合界面,而TACC3中的一个独特位点将ch-TOG招募到有丝分裂纺锤体中。转化酸性卷曲蛋白3 (TACC3)、结肠和肝脏肿瘤过表达基因(ch-TOG)和网格蛋白的复合物参与有丝分裂纺锤体组装和通过交联微管稳定着丝点纤维。目前还不清楚这种复合物是如何结合微管的,以及复合物中的蛋白质是如何相互作用的。TACC3和网格蛋白都被认为是纺锤体招募因子。我们绘制了复合体内的相互作用图,并表明TACC3和网格蛋白在纺锤体招募中是相互依赖的,必须相互作用才能将其中任何一个招募到纺锤体。网格蛋白的n端结构域与TACC3的TACC结构域串联形成微管相互作用表面,由TACC3 -网格蛋白结合协调。二亮氨酸基序和Aurora A磷酸化的TACC3上的丝氨酸558结合到网格蛋白的“脚踝”上。复合物内的另一个相互作用涉及TACC3卷曲线圈的停顿和ch-TOG中提出的新的第六个TOG结构域,这是ch-TOG微管定位所必需的,而不是TACC3 -网格蛋白。
Aurora A phosphorylation-induced interaction of TACC3 and clathrin coordinates adjacent domains in each protein to create a microtubule-binding interface, whereas a distinct site in TACC3 recruits ch-TOG to mitotic spindles. Acomplex of transforming acidic coiled-coil protein 3 (TACC3), colonic and hepatic tumor overexpressed gene (ch-TOG), and clathrin has been implicated in mitotic spindle assembly and in the stabilization of kinetochore fibers by cross-linking microtubules. It is unclear how this complex binds microtubules and how the proteins in the complex interact with one another. TACC3 and clathrin have each been proposed to be the spindle recruitment factor. We have mapped the interactions within the complex and show that TACC3 and clathrin were interdependent for spindle recruitment, having to interact in order for either to be recruited to the spindle. The N-terminal domain of clathrin and the TACC domain of TACC3 in tandem made a microtubule interaction surface, coordinated by TACC3–clathrin binding. A dileucine motif and Aurora A–phosphorylated serine 558 on TACC3 bound to the “ankle” of clathrin. The other interaction within the complex involved a stutter in the TACC3 coiled-coil and a proposed novel sixth TOG domain in ch-TOG, which was required for microtubule localization of ch-TOG but not TACC3–clathrin.
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