Biochemical analysis of PIFTC3, the Trypanosoma brucei orthologue of nematode DYF-13, reveals interactions with established and putative intraflagellar transport components.
Biochemical analysis of PIFTC3, the Trypanosoma brucei orthologue of nematode DYF-13, reveals interactions with established and putative intraflagellar transport components.
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DOI：
10.1111/j.1365-2958.2010.07322.x
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发表时间：
2010-10
影响因子：
3.6
通讯作者：
Ullu E
中科院分区：
文献类型：
--
作者：
Franklin JB;Ullu E
DYF-13最初在C.在染料填充化学感受突变体的集合中的秀丽线虫，是几种被分类为参与鞭毛内转运（IFT），蛋白复合物沿着纤毛和鞭毛的双向运动，特别是在顺行IFT中的蛋白质之一。虽然遗传学研究已经强调了DYF-13在线虫感觉纤毛和锥虫鞭毛生物发生中的基本作用，但DYF-13的生化研究却相对滞后。在这里，我们表明，在布氏锥虫的直系同源DYF 13，PIFC 3，参与了一个大分子复合物约660 kDa。亲和纯化的PIFC 3的质谱分析揭示了IFT复合物B的几种组分以及推定的IFT因子DYF-1、DYF-3、DYF-11/Elipsa和IFTA-2的直向同源物。DYF-11被进一步分析，并显示集中在基体附近和鞭毛中，并且是鞭毛伸长所需的。此外，通过共免疫沉淀，我们检测到DYF-13和IFT复合物A的一个组分IFT 122之间的相互作用，这是逆行转运所需的。因此，我们的生化分析支持了C. elegans，DYF-13的锥虫直系同源物在IFT机制中起着核心作用。
DYF-13, originally identified in C. elegans within a collection of dye-filling chemosensory mutants, is one of several proteins that have been classified as putatively involved in intraflagellar transport (IFT), the bidirectional movement of protein complexes along cilia and flagella, and specifically in anterograde IFT. Although genetic studies have highlighted a fundamental role of DYF-13 in nematode sensory cilium and trypanosome flagellum biogenesis, biochemical studies on DYF-13 have lagged behind. Here, we show that in Trypanosoma brucei the ortholog to DYF13, PIFTC3, participates in a macromolecular complex of approximately 660 kDa. Mass spectroscopy of affinity purified PIFTC3 revealed several components of IFT complex B as well as orthologs of putative IFT factors DYF-1, DYF-3, DYF-11/Elipsa, and IFTA-2. DYF-11 was further analyzed and shown to be concentrated near the basal bodies and in the flagellum, and to be required for flagellum elongation. In addition, by coimunoprecipitation we detected an interaction between DYF-13 and IFT122, a component of IFT complex A, which is required for retrograde transport. Thus, our biochemical analysis supports the model, proposed by genetic analysis in C. elegans, that the trypanosome ortholog of DYF-13 plays a central role in the IFT mechanism.
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小鼠IFT复合物的表征B。
影响因子：
--
作者：
Follit, John A.;Xu, Fenghui;Keady, Brian T.;Pazour, Gregory J.
通讯作者：
Pazour, Gregory J.
影响因子：
11.4
作者：
Kohl, L;Robinson, D;Bastin, P
通讯作者：
Bastin, P
影响因子：
3.3
作者：
Efimenko, Evgeni;Blacque, Oliver E.;Swoboda, Peter
通讯作者：
Swoboda, Peter
影响因子：
4.5
作者：
Li, Chunmei;Inglis, Peter N.;Leitch, Carmen C.;Efimenko, Evgeni;Zaghloul, Norann A.;Mok, Calvin A.;Davis, Erica E.;Bialas, Nathan J.;Healey, Michael P.;Heon, Elise;Zhen, Mei;Swoboda, Peter;Katsanis, Nicholas;Leroux, Michel R.
通讯作者：
Leroux, Michel R.
IFT驱动素的功能调节扩展了秀丽隐杆线虫中纤毛神经元的感觉曲目。
影响因子：
7.8
作者：
Evans, JE;Snow, JJ;Gunnarson, AL;Ou, GS;Stahlberg, H;McDonald, KL;Scholey, JM
通讯作者：
Scholey, JM