Chien S, Chung C Y, Sukumaran S, Osborne N, Lee S, Ellsworth C, McNally J G, Firtel R A
Section of Cell and Developmental Biology, Division of Biology, Center for Molecular Genetics, University of California, San Diego, USA.
Mol Biol Cell. 2000 Apr;11(4):1275-91. doi: 10.1091/mbc.11.4.1275.
We have identified limB, a gene encoding a novel LIM domain-containing protein, LIM2, in a screen for genes required for morphogenesis. limB null cells aggregate, although poorly, but they are unable to undergo morphogenesis, and the aggregates arrest at the mound stage. limB null cells exhibit an aberrant actin cytoskeleton and have numerous F-actin-enriched microspikes. The cells exhibit poor adhesion to a substratum and do not form tight cell-cell agglomerates in suspension. Furthermore, limB null cells are unable to properly polarize in chemoattractant gradients and move very poorly. Expression of limB from a prestalk-specific but not a prespore-specific promoter complements the morphogenetic defects of the limB null strain, suggesting that the limB null cell developmental defect results from an inability to properly sort prestalk cells. LIM2 protein is enriched in the cortex of wild-type cells, although it does not colocalize with the actin cytoskeleton. Our analysis indicates that LIM2 is a new regulatory protein that functions to control rearrangements of the actin cytoskeleton and is required for cell motility and chemotaxis. Our findings may be generally applicable to understanding pathways that control cell movement and morphogenesis in all multicellular organisms. Structure function studies on the LIM domains are presented.
我们在筛选形态发生所需基因的过程中,鉴定出了limB基因,它编码一种新型含LIM结构域的蛋白质LIM2。limB基因缺失的细胞能够聚集,尽管聚集效果不佳,但它们无法进行形态发生,并且聚集体在丘状阶段停滞。limB基因缺失的细胞表现出异常的肌动蛋白细胞骨架,并有许多富含F-肌动蛋白的微刺。这些细胞对基质的粘附性较差,在悬浮液中不能形成紧密的细胞间团聚体。此外,limB基因缺失的细胞在趋化因子梯度中无法正确极化,移动能力也很差。从柄细胞特异性而非孢子前体细胞特异性启动子表达limB可弥补limB基因缺失菌株的形态发生缺陷,这表明limB基因缺失细胞的发育缺陷是由于无法正确分选柄细胞所致。LIM2蛋白在野生型细胞的皮层中富集,尽管它与肌动蛋白细胞骨架不共定位。我们的分析表明,LIM2是一种新的调节蛋白,其功能是控制肌动蛋白细胞骨架的重排,是细胞运动和趋化作用所必需的。我们的发现可能普遍适用于理解控制所有多细胞生物中细胞运动和形态发生的途径。本文还介绍了对LIM结构域的结构功能研究。