Published online 8 March 2005
Article |
Expression of complementary RNA from chloroplast transgenes affects editing efficiency of transgene and endogenous chloroplast transcripts
Department of Molecular Biology and Genetics, Cornell University Ithaca, NY 14853, USA 1Department of Plant Biology, Cornell University Ithaca, NY 14853, USA
*To whom correspondence should be addressed. Tel: +1 607 254 4833; Fax: +1 607 255 6249; Email: mrh5{at}cornell.edu
Received December 19, 2004. Revised February 17, 2005. Accepted February 17, 2005.
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.
CiteULike 
Connotea 
Del.icio.us What's this?
This article has been cited by other articles:
|
|
 |
|
  M. L. Hayes, M. L. Reed, C. E. Hegeman, and M. R. Hanson Sequence elements critical for efficient RNA editing of a tobacco chloroplast transcript in vivo and in vitro Nucleic Acids Res., August 7, 2006; 34(13): 3742 - 3754. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 S. Holec, H. Lange, K. Kuhn, M. Alioua, T. Borner, and D. Gagliardi Relaxed transcription in Arabidopsis mitochondria is counterbalanced by RNA stability control mediated by polyadenylation and polynucleotide phosphorylase. Mol. Cell. Biol., April 1, 2006; 26(7): 2869 - 2876. [Abstract] [Full Text] [PDF]
|
|