Details
| Original language | English |
|---|---|
| Pages (from-to) | 357-67 |
| Number of pages | 11 |
| Journal | Plant molecular biology |
| Volume | 88 |
| Issue number | 4-5 |
| Publication status | Published - Jul 2015 |
Abstract
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.
Keywords
- Arabidopsis/genetics, Arabidopsis Proteins/genetics, Base Sequence, Binding Sites/genetics, Chloroplasts/genetics, DNA, Intergenic, Endoribonucleases/genetics, Gene Expression, Genes, Plant, Molecular Sequence Data, Mutation, Phosphoproteins/genetics, Photosystem II Protein Complex/genetics, Plants, Genetically Modified, RNA Precursors/genetics, RNA Processing, Post-Transcriptional, RNA, Antisense/genetics, RNA, Plant/genetics
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In: Plant molecular biology, Vol. 88, No. 4-5, 07.2015, p. 357-67.
Research output: Contribution to journal › Article › Research › peer review
}
TY - JOUR
T1 - 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
AU - Chevalier, Fabien
AU - Ghulam, Mustafa Malik
AU - Rondet, Damien
AU - Pfannschmidt, Thomas
AU - Merendino, Livia
AU - Lerbs-Mache, Silva
PY - 2015/7
Y1 - 2015/7
N2 - 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.
AB - 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.
KW - Arabidopsis/genetics
KW - Arabidopsis Proteins/genetics
KW - Base Sequence
KW - Binding Sites/genetics
KW - Chloroplasts/genetics
KW - DNA, Intergenic
KW - Endoribonucleases/genetics
KW - Gene Expression
KW - Genes, Plant
KW - Molecular Sequence Data
KW - Mutation
KW - Phosphoproteins/genetics
KW - Photosystem II Protein Complex/genetics
KW - Plants, Genetically Modified
KW - RNA Precursors/genetics
KW - RNA Processing, Post-Transcriptional
KW - RNA, Antisense/genetics
KW - RNA, Plant/genetics
U2 - 10.1007/s11103-015-0325-y
DO - 10.1007/s11103-015-0325-y
M3 - Article
C2 - 26012647
VL - 88
SP - 357
EP - 367
JO - Plant molecular biology
JF - Plant molecular biology
SN - 0167-4412
IS - 4-5
ER -