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环磷酸腺苷依赖性蛋白激酶在前孢子细胞中的表达足以诱导盘基网柄菌中的孢子细胞分化。

Expression of cAMP-dependent protein kinase in prespore cells is sufficient to induce spore cell differentiation in Dictyostelium.

作者信息

Mann S K, Richardson D L, Lee S, Kimmel A R, Firtel R A

机构信息

Department of Biology, University of California, San Diego, La Jolla 92093-0634.

出版信息

Proc Natl Acad Sci U S A. 1994 Oct 25;91(22):10561-5. doi: 10.1073/pnas.91.22.10561.

Abstract

The activity of cAMP-dependent protein kinase (PKA) is required for proper development at several stages during the Dictyostelium life cycle. We present evidence that activation of PKA is rate-limiting for the differentiation of prespore cells to spores and that PKA activation may be the developmental trigger for sporulation. Strains that overexpress the gene encoding the catalytic subunit of PKA (PKAcat) or lack a functional regulatory subunit (rdeC strains) undergo rapid, heterochronic development. We show that overexpression of PKAcat in prespore cell is sufficient to directly induce expression of the spore maturation marker spiA and differentiation to spores, in a cell-autonomous manner. Moreover, overexpression of PKAcat in prespore cells can bypass a mutation that blocks an earlier developmental step to induce spiA expression. Our results suggest that the regulatory pathway in prespore cells between the activation of PKA and spiA induction/spore maturation is quite short and that PKAcat expression in prespore cells may mediate spore differentiation at the level of transcription. This induction of sporulation requires the prior activation of the prespore cell pathway. In addition, we show that beta-galactosidase activity expressed from a PKAcat promoter/lacZ reporter construct is highly enriched in the anterior prestalk A region during the tipped aggregate, slug, and early culminant stages and that this pattern switches abruptly to a prespore pattern at the time of spore maturation, supporting the proposed role of PKA in this process.

摘要

环磷酸腺苷依赖性蛋白激酶(PKA)的活性在盘基网柄菌生命周期的几个阶段对于正常发育是必需的。我们提供的证据表明,PKA的激活是前孢子细胞分化为孢子的限速因素,并且PKA激活可能是孢子形成的发育触发因素。过表达编码PKA催化亚基的基因(PKAcat)或缺乏功能性调节亚基的菌株(rdeC菌株)会经历快速的异时发育。我们表明,在前孢子细胞中过表达PKAcat足以以细胞自主的方式直接诱导孢子成熟标记物spiA的表达并分化为孢子。此外,在前孢子细胞中过表达PKAcat可以绕过一个阻断早期发育步骤的突变来诱导spiA表达。我们的结果表明,前孢子细胞中PKA激活与spiA诱导/孢子成熟之间的调节途径相当短,并且前孢子细胞中PKAcat的表达可能在转录水平介导孢子分化。这种孢子形成的诱导需要前孢子细胞途径的预先激活。此外,我们表明,从PKAcat启动子/ lacZ报告构建体表达的β-半乳糖苷酶活性在倾斜聚集体、蛞蝓和早期子实体阶段在前柄A区域高度富集,并且这种模式在孢子成熟时突然转变为前孢子模式,支持了PKA在此过程中的拟议作用。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f547/45061/1aa2c468fbaf/pnas01144-0329-a.jpg

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