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PMID:20561259

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Citation

Nishimura, K, Ashida, H, Ogawa, T and Yokota, A (2010) A DEAD box protein is required for formation of a hidden break in Arabidopsis chloroplast 23S rRNA. Plant J. 63:766-77

Abstract

In plant chloroplasts, the ribosomal RNA (rRNA) of the large subunit of the ribosome undergoes post-maturation fragmentation processing. This processing consists of site-specific cleavage that generates gapped, discontinuous rRNA molecules. However, the molecular mechanism underlying introduction of the gap structure (the 'hidden break') is poorly understood. Here, we found that the DEAD box protein RH39 plays a key role in introduction of the hidden break into the 23S rRNA in Arabidopsis chloroplasts. Genetic screening for an Arabidopsis plant with a drastically reduced level of ribulose-1,5-bisphosphate carboxylase/oxygenase identified an RH39 mutant. The levels of other chloroplast-encoded photosynthetic proteins were also severely reduced. The reductions were not due to a failure of transcription, but rather inefficiency in translation. RNA gel blotting revealed incomplete fragmentation of 23S rRNA in chloroplasts during maturation. In vitro analysis with recombinant RH39 suggested that the protein binds to the adjacent sequence upstream of the hidden break site to exert its function. We propose a molecular mechanism for the RH39-mediated fragmentation processing of 23S rRNA in chloroplasts.

Links

PubMed Online version:10.1111/j.1365-313X.2010.04276.x

Keywords

Amino Acid Sequence; Arabidopsis/genetics; Arabidopsis/metabolism; Arabidopsis Proteins/genetics; Arabidopsis Proteins/metabolism; Blotting, Northern; Blotting, Western; Chloroplasts/metabolism; DEAD-box RNA Helicases/genetics; DEAD-box RNA Helicases/metabolism; Genes, Essential/genetics; Green Fluorescent Proteins/genetics; Green Fluorescent Proteins/metabolism; Molecular Sequence Data; Mutation; Protein Binding; RNA, Chloroplast/genetics; RNA, Chloroplast/metabolism; RNA, Ribosomal, 23S/genetics; RNA, Ribosomal, 23S/metabolism; Recombinant Proteins/metabolism; Ribulose-Bisphosphate Carboxylase/genetics; Ribulose-Bisphosphate Carboxylase/metabolism; Sequence Homology, Amino Acid

Significance

Annotations

Gene product Qualifier GO Term Evidence Code with/from Aspect Extension Notes Status

ARATH:RH39

enables

GO:0016887: ATP hydrolysis activity

ECO:0000314: direct assay evidence used in manual assertion

F

Seeded From UniProt

complete

ARATH:RH39

acts_upstream_of_or_within

GO:0110102: ribulose bisphosphate carboxylase complex assembly

ECO:0000315: mutant phenotype evidence used in manual assertion

P

Seeded From UniProt

complete

ARATH:RH39

enables

GO:0070180: large ribosomal subunit rRNA binding

ECO:0000314: direct assay evidence used in manual assertion

F

Seeded From UniProt

complete

ARATH:RH39

acts_upstream_of_or_within

GO:1901259: chloroplast rRNA processing

ECO:0000315: mutant phenotype evidence used in manual assertion

P

Seeded From UniProt

complete


See also

References

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