Coincident sequence-specific RNA degradation of linked transgenes in the plant genome
Identifieur interne : 002577 ( Main/Exploration ); précédent : 002576; suivant : 002578Coincident sequence-specific RNA degradation of linked transgenes in the plant genome
Auteurs : Megumi Kasai [Japon] ; Maiko Koseki [Japon] ; Kazunori Goto [Japon] ; Chikara Masuta [Japon] ; Shiho Ishii [Japon] ; Roger P. Hellens [Nouvelle-Zélande] ; Akito Taneda [Japon] ; Akira Kanazawa [Japon]Source :
- Plant Molecular Biology [ 0167-4412 ] ; 2012-02-01.
English descriptors
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Abstract
Abstract: The expression of transgenes in plant genomes can be inhibited by either transcriptional gene silencing or posttranscriptional gene silencing (PTGS). Overexpression of the chalcone synthase-A (CHS-A) transgene triggers PTGS of CHS-A and thus results in loss of flower pigmentation in petunia. We previously demonstrated that epigenetic inactivation of CHS-A transgene transcription leads to a reversion of the PTGS phenotype. Although neomycin phosphotransferase II (nptII), a marker gene co-introduced into the genome with the CHS-A transgene, is not normally silenced in petunia, even when CHS-A is silenced, here we found that nptII was silenced in a petunia line in which CHS-A PTGS was induced, but not in the revertant plants that had no PTGS of CHS-A. Transcriptional activity, accumulation of short interfering RNAs, and restoration of mRNA level after infection with viruses that had suppressor proteins of gene silencing indicated that the mechanism for nptII silencing was posttranscriptional. Read-through transcripts of the CHS-A gene toward the nptII gene were detected. Deep-sequencing analysis revealed a striking difference between the predominant size class of small RNAs produced from the read-through transcripts (22 nt) and that from the CHS-A RNAs (21 nt). These results implicate the involvement of read-through transcription and distinct phases of RNA degradation in the coincident PTGS of linked transgenes and provide new insights into the destabilization of transgene expression.
Url:
DOI: 10.1007/s11103-011-9863-0
Affiliations:
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Overexpression of the chalcone synthase-A (CHS-A) transgene triggers PTGS of CHS-A and thus results in loss of flower pigmentation in petunia. We previously demonstrated that epigenetic inactivation of CHS-A transgene transcription leads to a reversion of the PTGS phenotype. Although neomycin phosphotransferase II (nptII), a marker gene co-introduced into the genome with the CHS-A transgene, is not normally silenced in petunia, even when CHS-A is silenced, here we found that nptII was silenced in a petunia line in which CHS-A PTGS was induced, but not in the revertant plants that had no PTGS of CHS-A. Transcriptional activity, accumulation of short interfering RNAs, and restoration of mRNA level after infection with viruses that had suppressor proteins of gene silencing indicated that the mechanism for nptII silencing was posttranscriptional. Read-through transcripts of the CHS-A gene toward the nptII gene were detected. Deep-sequencing analysis revealed a striking difference between the predominant size class of small RNAs produced from the read-through transcripts (22 nt) and that from the CHS-A RNAs (21 nt). These results implicate the involvement of read-through transcription and distinct phases of RNA degradation in the coincident PTGS of linked transgenes and provide new insights into the destabilization of transgene expression.</div></front></TEI><affiliations><list><country><li>Japon</li><li>Nouvelle-Zélande</li></country></list><tree><country name="Japon"><noRegion><name sortKey="Kasai, Megumi" sort="Kasai, Megumi" uniqKey="Kasai M" first="Megumi" last="Kasai">Megumi Kasai</name></noRegion><name sortKey="Goto, Kazunori" sort="Goto, Kazunori" uniqKey="Goto K" first="Kazunori" last="Goto">Kazunori Goto</name><name sortKey="Ishii, Shiho" sort="Ishii, Shiho" uniqKey="Ishii S" first="Shiho" last="Ishii">Shiho Ishii</name><name sortKey="Kanazawa, Akira" sort="Kanazawa, Akira" uniqKey="Kanazawa A" first="Akira" last="Kanazawa">Akira Kanazawa</name><name sortKey="Kanazawa, Akira" sort="Kanazawa, Akira" uniqKey="Kanazawa A" first="Akira" last="Kanazawa">Akira Kanazawa</name><name sortKey="Koseki, Maiko" sort="Koseki, Maiko" uniqKey="Koseki M" first="Maiko" last="Koseki">Maiko Koseki</name><name sortKey="Masuta, Chikara" sort="Masuta, Chikara" uniqKey="Masuta C" first="Chikara" last="Masuta">Chikara Masuta</name><name sortKey="Taneda, Akito" sort="Taneda, Akito" uniqKey="Taneda A" first="Akito" last="Taneda">Akito Taneda</name></country><country name="Nouvelle-Zélande"><noRegion><name sortKey="Hellens, Roger P" sort="Hellens, Roger P" uniqKey="Hellens R" first="Roger P." last="Hellens">Roger P. Hellens</name></noRegion></country></tree></affiliations></record>Pour manipuler ce document sous Unix (Dilib)
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