Complex RNA metabolism in the chloroplast: an update on the psbB operon

Stoppel, Rhea; Meurer, Jörg

doi:10.1007/s00425-012-1782-z

Cited by 45 publications

(36 citation statements)

References 63 publications

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“…PsbN is expressed from the strand opposite the psbH gene in most cyanobacteria and in the psbB gene cluster in plastids of all plant lineages (Stoppel and Meurer, 2013). PsbN is one of the few plastid genes that has never been transferred successfully to the nucleus in photosynthetic eukaryotes during endosymbiosis (Race et al, 1999) presumably because its plastidic expression is coupled to early PSII assembly.…”

Section: Complementation Of the δPsbn Mutants By Ectopic Expression Omentioning

confidence: 99%

PsbN Is Required for Assembly of the Photosystem II Reaction Center in Nicotiana tabacum

et al. 2014

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ORCID ID: 0000-0003-2973-9514 (J.M.)The chloroplast-encoded low molecular weight protein PsbN is annotated as a photosystem II (PSII) subunit. To elucidate the localization and function of PsbN, encoded on the opposite strand to the psbB gene cluster, we raised antibodies and inserted a resistance cassette into PsbN in both directions. Both homoplastomic tobacco (Nicotiana tabacum) mutants ΔpsbN-F and ΔpsbN-R show essentially the same PSII deficiencies. The mutants are extremely light sensitive and failed to recover from photoinhibition. Although synthesis of PSII proteins was not altered significantly, both mutants accumulated only ;25% of PSII proteins compared with the wild type. Assembly of PSII precomplexes occurred at normal rates, but heterodimeric PSII reaction centers (RCs) and higher order PSII assemblies were not formed efficiently in the mutants. The ΔpsbN-R mutant was complemented by allotopic expression of the PsbN gene fused to the sequence of a chloroplast transit peptide in the nuclear genome. PsbN represents a bitopic trans-membrane peptide localized in stroma lamellae with its highly conserved C terminus exposed to the stroma. Significant amounts of PsbN were already present in dark-grown seedling. Our data prove that PsbN is not a constituent subunit of PSII but is required for repair from photoinhibition and efficient assembly of the PSII RC.

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Section: Complementation Of the δPsbn Mutants By Ectopic Expression Omentioning

confidence: 99%

PsbN Is Required for Assembly of the Photosystem II Reaction Center in Nicotiana tabacum

et al. 2014

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“…Legacy chloroplast RNA maturation analyses have used tedious gene by gene molecular techniques, restricting most detailed RNA analyses to several mono-or polycistronic transcripts including psbA, rbcL, atpI-atpH-atpF-atpA, and psbB-psbT-psbN-psbH-petB-petD (10), along with the ribosomal RNA operon, which together may not be fully representative of plastid RNA maturation pathways. Gene-by-gene analyses of processing factors may also have limited value.…”

Section: Introductionmentioning

confidence: 99%

Systematic sequencing of chloroplast transcript termini fromArabidopsis thalianareveals >200 transcription initiation sites and the extensive imprints of RNA-binding proteins and secondary structures

Castandet

Germain

Hotto³

et al. 2019

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Chloroplast transcription requires numerous quality control steps to generate the complex but selective mixture of accumulating RNAs. To gain insight into how this RNA diversity is achieved and regulated, we systematically mapped transcript ends by developing a protocol called Terminome-Seq. Using Arabidopsis thaliana as a model, we catalogued >215 primary 5' ends corresponding to transcription start sites (TSS), as well as 1,628 processed 5' ends and 1,299 3' ends. While most termini were found in intergenic regions, numerous abundant termini were also found within coding regions and introns, including several major TSS at unexpected locations. A consistent feature was the clustering of both 5' and 3' ends, contrasting with the prevailing description of discrete 5' termini, suggesting an imprecision of the transcription and/or RNA processing machinery. Numerous termini correlated with the extremities of small RNA footprints or predicted stem-loop structures, in agreement with the model of passive RNA protection. Terminome-Seq was also implemented for pnp1-1, a mutant lacking the processing enzyme polynucleotide phosphorylase. Nearly 2,000 termini were altered in pnp1-1, revealing a dominant role in shaping the transcriptome. In summary, Terminome-Seq permits precise delineation of the roles and regulation of the many factors involved in organellar transcriptome quality control.

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“…The resulting polycistronic psbB/T/H/petB/D transcript undergoes multiple splicing and processing events necessary for the formation of stable translatable mRNAs. The complex RNA metabolism of the psbB operon has recently been reviewed by Stoppel and Meurer (2013). Important processing events occur in the psbT-psbH intergenic region (Felder et al 2001;Zghidi-Abouzid et al 2011).…”

Section: Resultsmentioning

confidence: 99%

Characterization of the psbH precursor RNAs reveals a precise endoribonuclease cleavage site in the psbT/psbH intergenic region that is dependent on psbN gene expression

et al. 2015

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The plastid psbB operon harbours 5 genes, psbB, psbT, psbH, petB and petD. A sixth gene, the psbN gene, is located on the opposite DNA strand in the psbT/psbH intergenic region. Its transcription produces antisense RNA to a large part of the psbB pentacistronic mRNA. We have investigated whether transcription of the psbN gene, i.e. production of antisense RNA, influences psbT/psbH intergenic processing. Results reveal the existence of four different psbH precursor RNAs. Three of them result from processing and one is produced by transcription initiation. One of the processed RNAs is probably created by site-specific RNA cleavage. This RNA is absent in plants where the psbN gene is not transcribed suggesting that cleavage at this site is dependent on the formation of sense/antisense double-stranded RNA. In order to characterize the nuclease that might be responsible for double-stranded RNA cleavage, we analysed csp41a and csp41b knock-out mutants and the corresponding double mutant. Both CSP41 proteins are known to interact physically and CSP41a had been shown to cleave within 3'-untranslated region stem-loop structures, which contain double-stranded RNA, in vitro. We demonstrate that the psbH RNA, that is absent in plants where the psbN gene is not transcribed, is also strongly diminished in all csp41 plants. Altogether, results reveal a site-specific endoribonuclease cleavage event that seems to depend on antisense RNA and might implicate endoribonuclease activity of CSP41a.

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Complex RNA metabolism in the chloroplast: an update on the psbB operon

Cited by 45 publications

References 63 publications

PsbN Is Required for Assembly of the Photosystem II Reaction Center in Nicotiana tabacum

PsbN Is Required for Assembly of the Photosystem II Reaction Center in Nicotiana tabacum

Systematic sequencing of chloroplast transcript termini fromArabidopsis thalianareveals >200 transcription initiation sites and the extensive imprints of RNA-binding proteins and secondary structures

Characterization of the psbH precursor RNAs reveals a precise endoribonuclease cleavage site in the psbT/psbH intergenic region that is dependent on psbN gene expression

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