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在盘基网柄菌中鉴定依赖于STKA的基因。

Identification of STKA-dependent genes in Dictyostelium discoideum.

作者信息

Loughran G, Pinter K, Newell P C, Gross J D

机构信息

Dept. of Biochemistry, University of Oxford, United Kingdom.

出版信息

Differentiation. 2000 Oct;66(2-3):71-80. doi: 10.1046/j.1432-0436.2000.660202.x.

Abstract

During culmination of Dictyostelium aggregates, prespore and prestalk cells undergo terminal differentiation to form spores and a cellular stalk. Disruption of the cell-fate gene stkA leads to a phenotype in which all the cells destined to become spores end up as stalk cells. 'Stalky' mutants express normal levels of prespore cell transcripts but fail to produce the culmination-stage spore transcript spiA. The stkA gene encodes a putative GATA-type transcription factor (STKA). In order to identify possible downstream targets of STKA we used the technique of mRNA differential display and isolated four cDNA fragments that hybridise to mRNAs present during the later stages of development. All four gene tags were cloned and sequenced. mRNAs represented by these four sequence tags do not accumulate during culmination of 'stalky' cells and therefore must be specific to the spore pathway. By screening a cDNA library, longer cDNAs for all four were cloned and sequenced. Three of these contained complete protein-coding regions while only a partial cDNA was recovered for the fourth. One of the corresponding proteins has significant homology to a surface zinc metalloproteinase (GP63) of the protozoan parasite Leishmania, while another is closely related to a human pre-RNA binding protein (hnRNP R).

摘要

在盘基网柄菌聚集体发育成熟过程中,前孢子细胞和前柄细胞经历终末分化形成孢子和细胞性柄。细胞命运基因stkA的破坏导致一种表型,即所有注定成为孢子的细胞最终都变成了柄细胞。“多柄”突变体表达正常水平的前孢子细胞转录本,但无法产生发育成熟阶段的孢子转录本spiA。stkA基因编码一种假定的GATA型转录因子(STKA)。为了鉴定STKA可能的下游靶点,我们使用了mRNA差异显示技术,分离出了四个与发育后期出现的mRNA杂交的cDNA片段。所有四个基因标签都被克隆并测序。由这四个序列标签代表的mRNA在“多柄”细胞发育成熟过程中不积累,因此必定是孢子途径所特有的。通过筛选cDNA文库,克隆并测序了所有四个更长的cDNA。其中三个包含完整的蛋白质编码区,而第四个只获得了部分cDNA。其中一种相应的蛋白质与原生动物寄生虫利什曼原虫的表面锌金属蛋白酶(GP63)具有显著同源性,而另一种则与人类前体RNA结合蛋白(hnRNP R)密切相关。

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