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拟南芥磷脂酶C亚型的基因特异性表达与钙激活

Gene-specific expression and calcium activation of Arabidopsis thaliana phospholipase C isoforms.

作者信息

Hunt L, Otterhag L, Lee J C, Lasheen T, Hunt J, Seki M, Shinozaki K, Sommarin M, Gilmour D J, Pical C, Gray J E

机构信息

Department of Molecular Biology and Biotechnology, University of Sheffield, Sheffield S10 2TN, UK.

Department of Plant Biochemistry, Lund University PO Box 124, SE-221 00 Lund, Sweden.

出版信息

New Phytol. 2004 Jun;162(3):643-654. doi: 10.1111/j.1469-8137.2004.01069.x.

Abstract

•  PI-PLCs synthesise the calcium releasing second messenger IP . We investigated the expression patterns of the Arabidopsis PI-PLC gene family and measured in vitro activity of encoded enzymes. •  Gene specific RT-PCR and promoter-GUS fusions were used to analyse AtPLC gene expression patterns. The five available AtPLC cDNAs were expressed as fusion proteins in Escherichia coli. •  All members of the AtPLC gene family were expressed in multiple organs of the plant. AtPLC1, and AtPLC5 expression was localized to the vascular cells of roots and leaves with AtPLC5::GUS also detected in the guard cells. AtPLC4::GUS was detected in pollen and cells of the stigma surface. In seedlings, AtPLC2 and AtPLC3 were constitutively expressed, while AtPLCs 1, 4 and 5 were induced by abiotic stresses. AtPLC1-5 were all shown to have phospholipase C activity in the presence of calcium ions. •  AtPLCs showed limited tissue specific expression and expression of at least three genes was increased by abiotic stress. The differing calcium sensitivities of recombinant AtPLC protein activities may provide a mechanism for generating calcium signatures.

摘要

• 磷脂酰肌醇特异性磷脂酶C(PI-PLCs)合成释放钙的第二信使肌醇磷酸(IP)。我们研究了拟南芥PI-PLC基因家族的表达模式,并测定了编码酶的体外活性。

• 利用基因特异性逆转录聚合酶链反应(RT-PCR)和启动子-葡萄糖苷酶(GUS)融合技术分析AtPLC基因的表达模式。将五个可用的AtPLC cDNA在大肠杆菌中表达为融合蛋白。

• AtPLC基因家族的所有成员都在植物的多个器官中表达。AtPLC1和AtPLC5的表达定位于根和叶的维管细胞,AtPLC5::GUS在保卫细胞中也有检测到。AtPLC4::GUS在花粉和柱头表面的细胞中被检测到。在幼苗中,AtPLC2和AtPLC3组成型表达,而AtPLC1、4和5受非生物胁迫诱导。在钙离子存在的情况下,AtPLC1-5均显示具有磷脂酶C活性。

• AtPLCs表现出有限的组织特异性表达,并且至少三个基因的表达受非生物胁迫上调。重组AtPLC蛋白活性的不同钙敏感性可能为产生钙信号提供一种机制。

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