Cloning and characterization of glycine-rich RNA-binding protein cDNAs in the moss Physcomitrella patens
Cloning and characterization of glycine-rich RNA-binding protein cDNAs in the moss Physcomitrella patens
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DOI：
10.1093/pcp/pch005
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发表时间：
2004-01-01
影响因子：
4.9
通讯作者：
Sato, N
中科院分区：
文献类型：
--
作者：
Nomata, T;Kabeya, Y;Sato, N
我们从小立碗藓中分离出编码富含甘氨酸的RNA结合蛋白（GRP）的基因PpGRP 1、PpGRP 2和PpGRP 3的三个cDNA。从7日龄的扁胸蛙原丝体的poly（A）（+）RNA制备的cDNA文库中分离到3个全长cDNA克隆。PpGRP 1、PpGRP 2和PpGRP 3分别编码162、178和155个氨基酸残基的多肽。初步的基因组测序表明，PpGRP 3基因中的三个内含子的位置与拟南芥GRP基因中的内含子的位置相似。PpGRP 3具有一个假定的转运序列。PpGRP 1-sGFP和PpGRP 2-sGFP融合物靶向细胞核，而PpGRP 3-sGFP融合物靶向线粒体。这些PpGRP转录本的水平以及PpGRP蛋白的水平增加后，冷处理。同聚RNA分析显示PpGRP 3蛋白对poly（U）和poly（G）具有高亲和力。系统发育分析结果表明，在绿色植物进化的早期，GRP的核和线粒体形式已经确立。
We isolated three cDNAs for the genes PpGRP1, PpGRP2 and PpGRP3 that encode glycine-rich RNA-binding proteins (GRPs) from Physcomitrella patens. Three full-length cDNA clones were isolated from a cDNA library prepared from poly(A)(+) RNA from 7-day-old protonemata of P patens. They were named PpGRP1, PpGRP2 and PpGRP3, which encode putative polypeptides of 162, 178 and 155 residues, respectively. Preliminary genomic sequencing suggested that the positions of the three introns in the PpGRP3 gene are similar to those of introns in Arabidopsis GRP genes. PpGRP3 had a putative transit sequence. The PpGRP1-sGFP and PpGRP2-sGFP fusions were targeted to the cell nucleus, while PpGRP3-sGFP fusion was targeted to mitochondria. The level of these PpGRP transcripts as well as that of PpGRP proteins increased after cold treatment. Homoribopolymer RNA assay revealed that PpGRP3 protein show high affinity for poly(U) and poly(G). Results of phylogenetic analysis suggest that the nuclear and mitochondrial forms of GRP have been established early during the evolution of green plants.