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盘基网柄菌中对形态发生和前孢子细胞分化至关重要的一个基因的鉴定与分析。

Identification and analysis of a gene that is essential for morphogenesis and prespore cell differentiation in Dictyostelium.

作者信息

Yasukawa H, Mohanty S, Firtel R A

机构信息

Department of Biology, Center for Molecular Genetics, University of California, San Diego, La Jolla CA 92093-0634, USA.

出版信息

Development. 1998 Jul;125(14):2565-76. doi: 10.1242/dev.125.14.2565.

Abstract

We have identified a gene (PslA) that is expressed throughout Dictyostelium development and encodes a novel protein that is required for proper aggregation and subsequent cell-type differentiation and morphogenesis. pslA null (pslA-) cells produce large aggregation streams under conditions in which wild-type cells form discrete aggregates. Tips form along the stream, elongate to produce a finger, and eventually form a terminal structure that lacks a true sorus (spore head). More than half of the cells remain as a mass at the base of the developing fingers. The primary defect in the pslA- strain is the inability to induce prespore cell differentiation. Analyses of gene expression show a complete lack of prespore-specific gene expression and no mature spores are produced. In chimeras with wild-type cells, pslA- cells form the prestalk domain and normal, properly proportioned fruiting bodies can be produced. This indicates that pslA- cells are able to interact with wild-type cells and regulate patterning, even though pslA- cells are unable to express prespore cell-type-specific genes, do not participate in prespore cell differentiation and do not produce pslA- spores in the chimeras. While pslA- cells produce mature, vacuolated stalk cells during multicellular development, pslA- cells are unable to do so in vitro in response to exogenous DIF (a morphogen required for prestalk and stalk cell differentiation). These results indicate that pslA- cells exhibit a defect in the prestalk/stalk cell pathways under these experimental conditions. Our results suggest that PslA's primary function is to regulate prespore cell determination very early in the prespore pathway via a cell-autonomous mechanism, possibly at the time of the initial prestalk/prespore cell-fate decision. Indirect immunofluorescence of myc-tagged PslA localizes the protein to the nucleus, suggesting that PslA may function to control the prespore pathway at the level of transcription.

摘要

我们鉴定出了一个基因(PslA),它在盘基网柄菌的整个发育过程中都有表达,编码一种新型蛋白质,该蛋白质对于正常的聚集以及随后的细胞类型分化和形态发生是必需的。pslA基因敲除(pslA-)细胞在野生型细胞形成离散聚集体的条件下会产生大量聚集流。沿着流体会形成尖端,伸长形成手指状,最终形成缺乏真正孢子堆(孢子头)的终端结构。超过一半的细胞在发育中的手指状结构底部聚集成团。pslA-菌株的主要缺陷是无法诱导前孢子细胞分化。基因表达分析表明完全缺乏前孢子特异性基因表达,并且不产生成熟孢子。在与野生型细胞形成的嵌合体中,pslA-细胞形成前柄结构域,并且可以产生正常比例合适的子实体。这表明pslA-细胞能够与野生型细胞相互作用并调节模式形成,尽管pslA-细胞无法表达前孢子细胞类型特异性基因,不参与前孢子细胞分化,并且在嵌合体中不产生pslA-孢子。虽然pslA-细胞在多细胞发育过程中会产生成熟的、空泡化的柄细胞,但在体外,pslA-细胞对外源DIF(一种前柄和柄细胞分化所需的形态发生素)没有反应。这些结果表明,在这些实验条件下,pslA-细胞在前柄/柄细胞途径中存在缺陷。我们的结果表明,PslA的主要功能是通过细胞自主机制在孢子形成途径的早期非常早地调节前孢子细胞的决定,可能是在最初的前柄/前孢子细胞命运决定之时。对带有myc标签的PslA进行间接免疫荧光分析,将该蛋白质定位到细胞核,这表明PslA可能在转录水平上控制孢子形成途径。

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