期刊
PLANT AND CELL PHYSIOLOGY
卷 42, 期 10, 页码 1071-1078出版社
OXFORD UNIV PRESS
DOI: 10.1093/pcp/pce142
关键词
Arabidopsis thaliana; chloroplast; psbA gene; post-transcriptional; redox-regulated; RNA binding proteins
The 5 ' untranslated region (5 ' UTR) of the psbA mRNA (pshA encodes the PSII reaction center protein, DI) is a key site for FNA-protein interactions in the post-transcriptional regulation of gene expression. In this study, we mapped the major pshA mRNA 5 ' -terminus at -77 nt, and two minor termini clusters centered at -48 and -64 nt, upstream from the psbA translational start codon of Arabidopsis thaliana. RNA mobility shift, RNase protection and UV-crosslinking assays were used to characterize the interaction of chloroplast proteins with the RNA 5 ' UTR. RNA-protein interactions depended upon a thermolabile secondary structure and specific sequences in a 35 nt region of the 5 ' UTR, which were 80% conserved with the psbA 5 ' UTRs from five other plants. Major and minor proteins of 43- and 30-kDa, respectively, were detected by UV-crosslinking to RNA. Oxidizing conditions abolished the association of the proteins with the 5 ' UTR, while RNA-binding activity was recovered upon incubation with a reductant. Based on these findings, we hypothesize that post-transcriptional regulation of psbA gene expression in chloroplasts of vascular plants involves redox-dependent interactions between specific sequences in the 5 ' UTR and 43- and 30-kDa RNA-binding proteins.
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