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一种编码盘基网柄菌中前孢子细胞诱导因子的基因。

A gene encoding, prespore-cell-inducing factor in Dictyostelium discoideum.

作者信息

Kawata Takefumi, Nakagawa Manabu, Shimada Nao, Fujii Shigeru, Oohata Akiko A

机构信息

Department of Biology, Faculty of Science, Toho University, 2-2-1 Miyama, Funabashi, Chiba 274-8510, Japan.

出版信息

Dev Growth Differ. 2004 Aug;46(4):383-92. doi: 10.1111/j.1440-169x.2004.00749.x.

Abstract

Two factors that exist in conditioned medium (CM) of Dictyostelium discoideum induce amoebae to differentiate into prespore cells when they are incubated at a very low cell density in submerged monolayer culture. Previously, we purified one of them, a glycoprotein factor with an apparent molecular mass of 106 kDa, and we named it psi factor (psi, prespore-inducing factor). Based on the partial amino acid sequence of the purified psi factor, we have isolated the corresponding cDNA clone, which is expressed maximally at the loose mound stage. The cDNA encodes a novel protein and the predicted molecular mass of the mature secreted protein is 60 kDa. Knockout mutant strains of the psi factor gene, psiA(-), were created by targeted integration. Although these mutant strains appear to develop normally, CM from these mutants showed reduced prespore-cell-inducing activity. Rescuing the mutant strains by expression of psi factor under control of a constitutive promoter causes overproduction of psi factor protein and CM from such cells showed a 20-fold higher level of prespore-cell-inducing activity than that from wild-type cells. Further, CM from parental cells induced prespore cell division, while that from psiA null strains showed no cell division inducing activity. Our results indicate that psi factor protein is a novel type of growth factor that does not belong to any of the families of growth factor so far identified in animals.

摘要

盘基网柄菌(Dictyostelium discoideum)条件培养基(CM)中的两种因子,当在浸没单层培养中以非常低的细胞密度孵育时,可诱导变形虫分化为前孢子细胞。此前,我们纯化了其中一种,一种表观分子量为106 kDa的糖蛋白因子,并将其命名为psi因子(psi,前孢子诱导因子)。基于纯化的psi因子的部分氨基酸序列,我们分离出了相应的cDNA克隆,其在松散丘阶段表达量最高。该cDNA编码一种新型蛋白质,成熟分泌蛋白的预测分子量为60 kDa。通过靶向整合创建了psi因子基因的敲除突变株psiA(-)。尽管这些突变株似乎发育正常,但来自这些突变体的CM显示出前孢子细胞诱导活性降低。通过在组成型启动子控制下表达psi因子来拯救突变株,会导致psi因子蛋白过量产生,并且来自此类细胞的CM显示出比野生型细胞高20倍的前孢子细胞诱导活性。此外,来自亲代细胞的CM可诱导前孢子细胞分裂,而来自psiA缺失株的CM则没有细胞分裂诱导活性。我们的结果表明,psi因子蛋白是一种新型生长因子,不属于迄今为止在动物中鉴定出的任何生长因子家族。

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