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Expression of complementary RNA from chloroplast transgenes affects editing efficiency of transgene and endogenous chloroplast transcripts

The expression of angiosperm chloroplast genes is modified by C-to-U RNA editing. The mechanism for recognition of the ∼30 C targets of editing is not understood. There is no single consensus sequence surrounding editing sites, though sites can be grouped into small ‘clusters’ of two to five sites e...

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Autores principales: Hegeman, Carla E., Halter, Christine P., Owens, Thomas G., Hanson, Maureen R.
Formato: Texto
Lenguaje:English
Publicado: Oxford University Press 2005
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC1062874/
https://www.ncbi.nlm.nih.gov/pubmed/15755747
http://dx.doi.org/10.1093/nar/gki286
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author Hegeman, Carla E.
Halter, Christine P.
Owens, Thomas G.
Hanson, Maureen R.
author_facet Hegeman, Carla E.
Halter, Christine P.
Owens, Thomas G.
Hanson, Maureen R.
author_sort Hegeman, Carla E.
collection PubMed
description The expression of angiosperm chloroplast genes is modified by C-to-U RNA editing. The mechanism for recognition of the ∼30 C targets of editing is not understood. There is no single consensus sequence surrounding editing sites, though sites can be grouped into small ‘clusters’ of two to five sites exhibiting some sequence similarity. While complementary RNA that guides nucleotides for alteration has been detected in other RNA modification systems, it is not known whether complementary RNA is involved in chloroplast editing site recognition. We investigated the effect of expressing RNA antisense to the sequences −20 to +6 surrounding the RpoB-2 C target of editing, which is a member of a cluster that includes the PsbL-1 and Rps14-1 sites. Previous experiments had shown that chloroplast rpoB transgene transcripts carrying only these 27 nt were edited in vivo at the proper C. Though transcripts carrying sequences −31 to +60 surrounding the RpoB-2 sites were edited in chloroplast transgenic plants, transcripts carrying the −31 to +62 region followed by the 27 nt complementary region were not edited at all. In contrast, a similar construct, in which the C target as well as the preceding and subsequent nucleotides were mismatched within the 27 nt region, was efficiently edited. The presence of any of the four transgenes carrying RpoB-2 sequences in sense and/or antisense orientation resulted in reduced editing at the PsbL-1 site. Chloroplast transgenic plants expressing the three different antisense RNA constructs exhibited abnormal growth and development, though plants expressing the 92 nt sense transcripts were phenotypically normal.
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spelling pubmed-10628742005-03-08 Expression of complementary RNA from chloroplast transgenes affects editing efficiency of transgene and endogenous chloroplast transcripts Hegeman, Carla E. Halter, Christine P. Owens, Thomas G. Hanson, Maureen R. Nucleic Acids Res Article The expression of angiosperm chloroplast genes is modified by C-to-U RNA editing. The mechanism for recognition of the ∼30 C targets of editing is not understood. There is no single consensus sequence surrounding editing sites, though sites can be grouped into small ‘clusters’ of two to five sites exhibiting some sequence similarity. While complementary RNA that guides nucleotides for alteration has been detected in other RNA modification systems, it is not known whether complementary RNA is involved in chloroplast editing site recognition. We investigated the effect of expressing RNA antisense to the sequences −20 to +6 surrounding the RpoB-2 C target of editing, which is a member of a cluster that includes the PsbL-1 and Rps14-1 sites. Previous experiments had shown that chloroplast rpoB transgene transcripts carrying only these 27 nt were edited in vivo at the proper C. Though transcripts carrying sequences −31 to +60 surrounding the RpoB-2 sites were edited in chloroplast transgenic plants, transcripts carrying the −31 to +62 region followed by the 27 nt complementary region were not edited at all. In contrast, a similar construct, in which the C target as well as the preceding and subsequent nucleotides were mismatched within the 27 nt region, was efficiently edited. The presence of any of the four transgenes carrying RpoB-2 sequences in sense and/or antisense orientation resulted in reduced editing at the PsbL-1 site. Chloroplast transgenic plants expressing the three different antisense RNA constructs exhibited abnormal growth and development, though plants expressing the 92 nt sense transcripts were phenotypically normal. Oxford University Press 2005 2005-03-08 /pmc/articles/PMC1062874/ /pubmed/15755747 http://dx.doi.org/10.1093/nar/gki286 Text en © The Author 2005. Published by Oxford University Press. All rights reserved
spellingShingle Article
Hegeman, Carla E.
Halter, Christine P.
Owens, Thomas G.
Hanson, Maureen R.
Expression of complementary RNA from chloroplast transgenes affects editing efficiency of transgene and endogenous chloroplast transcripts
title Expression of complementary RNA from chloroplast transgenes affects editing efficiency of transgene and endogenous chloroplast transcripts
title_full Expression of complementary RNA from chloroplast transgenes affects editing efficiency of transgene and endogenous chloroplast transcripts
title_fullStr Expression of complementary RNA from chloroplast transgenes affects editing efficiency of transgene and endogenous chloroplast transcripts
title_full_unstemmed Expression of complementary RNA from chloroplast transgenes affects editing efficiency of transgene and endogenous chloroplast transcripts
title_short Expression of complementary RNA from chloroplast transgenes affects editing efficiency of transgene and endogenous chloroplast transcripts
title_sort expression of complementary rna from chloroplast transgenes affects editing efficiency of transgene and endogenous chloroplast transcripts
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC1062874/
https://www.ncbi.nlm.nih.gov/pubmed/15755747
http://dx.doi.org/10.1093/nar/gki286
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