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一种蛋白激酶基因的MAP/ERK家族同源物在矮牵牛的营养器官和雌性生殖器官中表达。

A homologue of the MAP/ERK family of protein kinase genes is expressed in vegetative and in female reproductive organs of Petunia hybrida.

作者信息

Decroocq-Ferrant V, Decroocq S, Van Went J, Schmidt E, Kreis M

机构信息

Université de Paris-Sud, IBP, URA-CNRS 1128, Biologie du Développement des Plantes, Orsay, France.

出版信息

Plant Mol Biol. 1995 Jan;27(2):339-50. doi: 10.1007/BF00020188.

Abstract

The mitogen activated protein (MAP) kinase pathway of eukaryotes is stimulated by many growth factors and is required for the integration of multiple cellular signals. In order to study the function of MAP kinases during plant ovule development we have synthesized a Petunia hybrida ovule-specific cDNA library and screened for MAP protein kinase-related sequences using a DNA probe obtained by PCR. A full-length cDNA clone was identified (PMEK for Petunia hybrida MAP/ERK-related protein kinase) and shown to encode a protein related to the family of MAP/ERK protein kinases. Southern blot analysis showed that PMEK is a member of a small multigene family in P. hybrida. The cDNA codes for a protein (PMEK1) of 44.4 kDa with an overall sequence identity of 44% to the products of the mammalian ERK/MAP kinase gene, and the budding yeast KSS1 and FUS3 genes. PMEK1 displays 96 and 80% identity respectively with the tobacco NTF3 and Arabidopsis ATMPK1 kinases, and only 50% to the more distantly related plant MAP kinase MsERK1 from alfalfa. The two phosphorylation sites found in the loop between subdomain VII and VIII in all the other MAP kinases are also present in PMEK1. RNA gel blot and RT-PCR analyses demonstrated that PMEK1 is expressed in vegetative organs and preferentially accumulated in female reproductive organs of P. hybrida. In situ hybridization experiments showed that in the reproductive organs PMEK1 is expressed only in the ovary and not in the stamen.

摘要

真核生物的丝裂原活化蛋白(MAP)激酶途径受多种生长因子刺激,是整合多种细胞信号所必需的。为了研究MAP激酶在植物胚珠发育过程中的功能,我们合成了矮牵牛胚珠特异性cDNA文库,并使用通过PCR获得的DNA探针筛选与MAP蛋白激酶相关的序列。鉴定出一个全长cDNA克隆(矮牵牛MAP/ERK相关蛋白激酶PMEK),并显示其编码一种与MAP/ERK蛋白激酶家族相关的蛋白质。Southern印迹分析表明,PMEK是矮牵牛中一个小的多基因家族的成员。该cDNA编码一种44.4 kDa的蛋白质(PMEK1),与哺乳动物ERK/MAP激酶基因、芽殖酵母KSS1和FUS3基因的产物的总体序列同一性为44%。PMEK1与烟草NTF3和拟南芥ATMPK1激酶的同一性分别为96%和80%,与苜蓿中亲缘关系较远的植物MAP激酶MsERK1的同一性仅为50%。在所有其他MAP激酶的亚结构域VII和VIII之间的环中发现的两个磷酸化位点在PMEK1中也存在。RNA凝胶印迹和RT-PCR分析表明,PMEK1在营养器官中表达,并在矮牵牛的雌性生殖器官中优先积累。原位杂交实验表明,在生殖器官中,PMEK1仅在子房表达,而不在雄蕊中表达。

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