Localization of MglA, an essential gliding motility protein in Myxococcus xanthus.
Localization of MglA, an essential gliding motility protein in Myxococcus xanthus.
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黄色粘球菌中一种重要的滑动运动蛋白 MglA 的定位。
DOI：
10.1002/cm.20447
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发表时间：
2010
期刊：
影响因子：
--
通讯作者：
Hartzell,PatriciaL
中科院分区：
文献类型：
--
作者：
Patryn,Jacek;Allen,Krisandra;Dziewanowska,Katarzyna;Otto,Rebecca;Hartzell,PatriciaL
MglA是一种与单体GTP酶相关的22 kDa蛋白，是黄色粘球菌A（冒险）和S（社会）运动的正常运作以及多细胞发育所必需的。为了确定MglA如何控制A运动性和S运动性，对MglA进行生物化学测定并确定其细胞位置。His标记的MglA在体外以几乎与Ras相同的速率缓慢水解GTP，表明MglA具有GTP酶活性。来自液体的固定细胞的免疫荧光显微镜显示MglA与跨越细胞长度的类似于MreB螺旋的螺旋轨道相关。MglA的分布模式取决于从其收获细胞的表面的类型。在1.5%（w/v）琼脂上滑动的细胞中，螺旋模式让位于在两极和沿着长轴的MglA-Yfp点状簇（侧向簇）。随着细胞前进，侧向簇出现在前极附近，与前极处的MglA-Yfp簇的强度降低一致。新形成的横向集群保持固定相对于基层的细胞向前移动，类似于局部粘附复合物描述的AglZ，蛋白伴侣的MglA。在甲基纤维素（一种在低细胞密度下刺激S-运动性的聚合物）中滑动的细胞中没有形成侧向簇;相反，MglA-Yfp在细胞质中扩散，并且在两极更集中。结果表明，有利于S运动的条件防止形成与A运动功能相关的MglA侧向簇。© 2010 Wiley利斯公司
MglA, a 22‐kDa protein related to monomeric GTPases, is required for the normal operation of the A (Adventurous) and S (Social) motility and for multicellular development ofMyxococcus xanthus. To determine how MglA controls A‐ and S‐motility, MglA was assayed biochemically and its cellular location was determined. His‐tagged MglA hydrolyzed GTP slowly in vitro at a rate nearly identical to that of Ras showing that MglA has GTPase activity. Immunofluorescence microscopy of fixed cells from liquid showed that MglA was associated with helical track similar to the MreB spiral that spanned the length of the cell. The distribution pattern of MglA depended on the type of surface from which cells were harvested. In cells gliding on 1.5% (w/v) agar, the helical pattern gave way to punctate clusters of MglA‐Yfp at the poles and along the long axis (lateral clusters). The lateral clusters emerged near the leading pole as the cell advanced coincident with a decrease in the intensity of the MglA‐Yfp cluster at the leading pole. Newly formed lateral clusters remained fixed with regard to the substratum as the cell moved forward, similar to focal adhesion complexes described for AglZ, a protein partner of MglA. Lateral clusters did not form in cells gliding in methylcellulose, a polymer that stimulates S‐motility at low cell density; rather MglA‐Yfp was diffuse in the cytoplasm and more concentrated at the poles. The results suggest that conditions that favor S‐motility prevent the formation of lateral clusters of MglA, which are associated with A‐motility functions. © 2010 Wiley‐Liss, Inc.
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