Tor2直接磷酸化AGC激酶Ypk2以调节肌动蛋白极化。

Tor2 directly phosphorylates the AGC kinase Ypk2 to regulate actin polarization.

作者信息

Kamada Yoshiaki, Fujioka Yuko, Suzuki Nobuo N, Inagaki Fuyuhiko, Wullschleger Stephan, Loewith Robbie, Hall Michael N, Ohsumi Yoshinori

机构信息

Department of Cell Biology, National Institute for Basic Biology, Maiodaiji-Cho, Okazaki, Japan.

出版信息

Mol Cell Biol. 2005 Aug;25(16):7239-48. doi: 10.1128/MCB.25.16.7239-7248.2005.

Abstract

The target of rapamycin (TOR) protein kinases, Tor1 and Tor2, form two distinct complexes (TOR complex 1 and 2) in the yeast Saccharomyces cerevisiae. TOR complex 2 (TORC2) contains Tor2 but not Tor1 and controls polarity of the actin cytoskeleton via the Rho1/Pkc1/MAPK cell integrity cascade. Substrates of TORC2 and how TORC2 regulates the cell integrity pathway are not well understood. Screening for multicopy suppressors of tor2, we obtained a plasmid expressing an N-terminally truncated Ypk2 protein kinase. This truncation appears to partially disrupt an autoinhibitory domain in Ypk2, and a point mutation in this region (Ypk2(D239A)) conferred upon full-length Ypk2 the ability to rescue growth of cells compromised in TORC2, but not TORC1, function. YPK2(D239A) also suppressed the lethality of tor2Delta cells, suggesting that Ypks play an essential role in TORC2 signaling. Ypk2 is phosphorylated directly by Tor2 in vitro, and Ypk2 activity is largely reduced in tor2Delta cells. In contrast, Ypk2(D239A) has increased and TOR2-independent activity in vivo. Thus, we propose that Ypk protein kinases are direct and essential targets of TORC2, coupling TORC2 to the cell integrity cascade.

摘要

雷帕霉素靶蛋白(TOR)激酶Tor1和Tor2在酿酒酵母中形成两种不同的复合物(TOR复合物1和2)。TOR复合物2(TORC2)包含Tor2但不包含Tor1,并通过Rho1/Pkc1/MAPK细胞完整性级联反应控制肌动蛋白细胞骨架的极性。TORC2的底物以及TORC2如何调节细胞完整性途径尚不清楚。通过筛选tor2的多拷贝抑制子,我们获得了一个表达N端截短的Ypk2蛋白激酶的质粒。这种截短似乎部分破坏了Ypk2中的自抑制结构域,并且该区域的一个点突变(Ypk2(D239A))赋予全长Ypk2挽救TORC2功能受损细胞生长的能力,但不能挽救TORC1功能受损细胞的生长。YPK2(D239A)也抑制了tor2Δ细胞的致死性,表明Ypks在TORC2信号传导中起重要作用。Ypk2在体外被Tor2直接磷酸化,并且在tor2Δ细胞中Ypk2活性大大降低。相反,Ypk2(D239A)在体内具有增加的且不依赖TOR2的活性。因此,我们提出Ypk蛋白激酶是TORC2的直接且重要的靶标,将TORC2与细胞完整性级联反应联系起来。

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