Direct interaction of Gas11 with microtubules: Implications for the dynein regulatory complex
Direct interaction of Gas11 with microtubules: Implications for the dynein regulatory complex
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DOI：
10.1002/cm.20196
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发表时间：
2007-06-01
影响因子：
--
通讯作者：
Crosbie, Rachelle H.
中科院分区：
文献类型：
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作者：
Bekker, Janine M.;Colantonio, Jessica R.;Crosbie, Rachelle H.
我们之前描述了与动力蛋白调节有关的细胞骨架相关蛋白的锥虫蛋白家族[Hill et al., J Biol Chem 2000; 275(50)：39369-39378； Hutchings 等人，《细胞生物学杂志》2002；156(5)：867-877； Rupp 和 Porter，《细胞生物学杂志》2003；162(1)：47-57]。来自布氏锥虫的锥虫蛋白是进化上保守的动力蛋白调节系统的一部分，该系统是调节鞭毛节拍所必需的。在莱茵衣藻中，锥虫蛋白同源物 (PF2) 是轴丝“动力蛋白调节复合物”(DRC) 的一部分，其功能是轴丝动力蛋白的可逆抑制剂 [Piperno 等人，J Cell Biol 1992；118(6):1455-1463； Gardner 等人，《细胞生物学杂志》1994；127(5)：1311-1325]。 DRC 由估计的七种与轴丝微管紧密相关的多肽组成。与轴丝的结合对于 DRC 功能至关重要，但它附着在微管晶格上的机制完全未知。我们证明，哺乳动物胰蛋白酶/PF2 同源物 Gas11 在体外和体内与微管相关。删除分析确定了一个新的微管结合域 (GMAD) 和一个减弱 Gas11-微管相互作用的独特区域 (IMAD)。使用单颗粒结合测定，我们证明 Gas 11 直接结合微管，并且 IMAD 减弱 GMAD 和微管之间的相互作用。 IMAD 能够与 GMAD 一起以顺式或反式方向发挥作用。 Gas 11 与微管直接连接的发现为了解动力蛋白调节复合物的结构特征提供了新的机制见解。细胞动力。细胞骨架 64:461-473, 2007。(C) 2007 Wiley-Liss, Inc.
We previously described the Trypanin family of cytoskeleton-associated proteins that have been implicated in dynein regulation [Hill et al., J Biol Chem 2000; 275(50):39369-39378; Hutchings et al., J Cell Biol 2002;156(5):867-877; Rupp and Porter, J Cell Biol 2003;162(1):47-57]. Trypanin from T. brucei is part of an evolutionarily conserved dynein regulatory system that is required for regulation of flagellar beat. In C. reinhardtii, the trypanin homologue (PF2) is part of an axonemal 'dynein regulatory complex' (DRC) that functions as a reversible inhibitor of axonemal dynein [Piperno et al., J Cell Biol 1992;118(6):1455-1463; Gardner et al., J Cell Biol 1994;127(5):1311-1325]. The DRC consists of an estimated seven polypeptides that are tightly associated with axonemal microtubules. Association with the axoneme is critical for DRC function, but the mechanism by which it attaches to the microtubule lattice is completely unknown. We demonstrate that Gas11, the mammalian trypanin/PF2 homologue, associates with microtubules in vitro and in vivo. Deletion analyses identified a novel microtubule-binding domain (GMAD) and a distinct region (IMAD) that attenuates Gas11-microtubule interactions. Using single-particle binding assays, we demonstrate that Gas 11 directly binds microtubules and that the IMAD attenuates the interaction between GMAD and the micrombule. IMAD is able to function in either a cis- or trans-orientation with GMAD. The discovery that Gas 11 provides a direct linkage to microtubules provides new mechanistic insight into the structural features of the dynein-regulatory complex. Cell Motil. Cytoskeleton 64:461-473, 2007. (C) 2007 Wiley-Liss, Inc.