Journal Article

Cloning and Characterization of Glycine-Rich RNA-Binding Protein cDNAs in the Moss <i>Physcomitrella patens</i>

Tsuyoshi Nomata, Yukihiro Kabeya and Naoki Sato

in Plant and Cell Physiology

Published on behalf of Japanese Society of Plant Physiologists

Volume 45, issue 1, pages 48-56
Published in print January 2004 | ISSN: 0032-0781
Published online January 2004 | e-ISSN: 1471-9053 | DOI: http://dx.doi.org/10.1093/pcp/pch005
Cloning and Characterization of Glycine-Rich RNA-Binding Protein cDNAs in the Moss Physcomitrella patens

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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.

Keywords: Keywords: Cold regulation — Glycine-rich protein — Mitochondrion — Physcomitrella patens — RNA-binding protein.; Abbreviations: CaMV, cauliflower mosaic virus; CBB, Coomassie brilliant blue; DIG, digoxigenin; GFP, green fluorescent protein; GRP, glycine-rich RNA-binding protein; ML, maximum likelihood; NJ, neighbor joining; RRM, RNA recognition motif; RNP, ribonucleoprotein; UTR, untranslated region.

Journal Article.  5465 words.  Illustrated.

Subjects: Biochemistry ; Molecular and Cell Biology ; Plant Sciences and Forestry

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