Identification of TOEFAZ1-interacting proteins reveals key regulators of Trypanosoma brucei cytokinesis.
Identification of TOEFAZ1-interacting proteins reveals key regulators of Trypanosoma brucei cytokinesis.
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DOI：
10.1111/mmi.13986
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发表时间：
2018-08
影响因子：
3.6
通讯作者：
de Graffenried CL
中科院分区：
文献类型：
--
作者：
Hilton NA;Sladewski TE;Perry JA;Pataki Z;Sinclair-Davis AN;Muniz RS;Tran HL;Wurster JI;Seo J;de Graffenried CL
原生生物寄生虫布氏锥虫是一种专性细胞外病原体，在细胞分裂过程中保留其高度极化的形态，并已发展出一种新的细胞动力学过程独立于非肌肉肌球蛋白II。Polo样激酶同系物TbPLK对于分裂期间细胞极性的传递和胞质分裂是必不可少的。我们以前确定了一个假定的TbPLK底物命名为尖端的延伸FAZ 1（TOEFAZ 1）作为一个必要的动质体特异性组成部分的T。布鲁氏菌细胞动力学机制我们使用TOEFAZ1进行了邻近依赖性生物素化（BioID）筛选，以鉴定参与胞质分裂的其他蛋白质。使用定量蛋白质组学方法，我们确定了近500个TOEFAZ1近端蛋白，并进一步详细描述了59个。在候选人中，我们确定了一个基本的假定磷酸酶，调节TOEFAZ 1和TbPLK，一个以前未知的蛋白质，这是一个新的细胞后部的组装所必需的表达水平和本地化，和微管加端定向孤儿驱动蛋白，这是完成切割沟内移所需的。这些蛋白的鉴定为T.布氏胞质分裂，并建立TOEFAZ 1作为这一重要的和复杂配置的过程中动质体的关键组成部分。布氏锥虫采用一种新的细胞动力学机制，该机制沿着细胞的长轴进行，并需要动质体特异性蛋白TOEFAZ 1。使用TOEFAZ 1作为诱饵的邻近生物素化已经鉴定了T.布鲁氏菌胞质分裂，包括驱动蛋白、磷酸酶和与膜下微管相关的蛋白质。这些组分说明了T.布氏胞质分裂，并将允许更好地理解这一过程的机制。
The protist parasite Trypanosoma brucei is an obligate extracellular pathogen that retains its highly-polarized morphology during cell division and has evolved a novel cytokinetic process independent of non-muscle myosin II. The polo-like kinase homolog TbPLK is essential for transmission of cell polarity during division and for cytokinesis. We previously identified a putative TbPLK substrate named Tip of the Extending FAZ 1 (TOEFAZ1) as an essential kinetoplastid-specific component of the T. brucei cytokinetic machinery. We performed a proximity-dependent biotinylation (BioID) screen using TOEFAZ1 as a means to identify additional proteins that are involved in cytokinesis. Using quantitative proteomic methods, we identified nearly 500 TOEFAZ1-proximal proteins and characterized 59 in further detail. Among the candidates, we identified an essential putative phosphatase that regulates the expression level and localization of both TOEFAZ1 and TbPLK, a previously uncharacterized protein that is necessary for the assembly of a new cell posterior, and a microtubule plus-end directed orphan kinesin that is required for completing cleavage furrow ingression. The identification of these proteins provides new insight into T. brucei cytokinesis and establishes TOEFAZ1 as a key component of this essential and uniquely-configured process in kinetoplastids. Trypanosoma brucei employs a novel cytokinetic mechanism that proceeds along the long axis of the cell and requires the kinetoplastid-specific protein TOEFAZ1. Proximity biotinylation using TOEFAZ1 as the bait has identified novel and essential proteins involved in T. brucei cytokinesis, including a kinesin, a phosphatase, and a protein associated with the subpellicular microtubules. These components illustrate the unique configuration of T. brucei cytokinesis and will allow a better mechanistic understanding of this process.
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影响因子：
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