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Titolo:
Altering the 3 ' UTR endonucleolytic cleavage site of a Chlamydomonas chloroplast mRNA affects 3 '-end maturation in vitro but not in vivo
Autore:
Rott, R; Liveanu, V; Drager, RG; Higgs, D; Stern, DB; Schuster, G;
Indirizzi:
Technion Israel Inst Technol, Dept Biol, IL-32000 Haifa, Israel Technion Israel Inst Technol Haifa Israel IL-32000 L-32000 Haifa, Israel Cornell Univ, Boyce Thompson Inst Plant Res, Ithaca, NY 14853 USA Cornell Univ Ithaca NY USA 14853 son Inst Plant Res, Ithaca, NY 14853 USA
Titolo Testata:
PLANT MOLECULAR BIOLOGY
fascicolo: 4, volume: 40, anno: 1999,
pagine: 679 - 686
SICI:
0167-4412(1999)40:4<679:AT3'UE>2.0.ZU;2-M
Fonte:
ISI
Lingua:
ENG
Soggetto:
RNA-BINDING PROTEIN; MESSENGER-RNA; SPINACH CHLOROPLAST; INVERTED REPEATS; END MATURATION; ATPB GENE; DEGRADATION; REINHARDTII; COMPLEX; INVIVO;
Keywords:
atpB; Chlamydomonas reinhardtii; chloroplast gene expression; mRNA processing;
Tipo documento:
Article
Natura:
Periodico
Settore Disciplinare:
Agriculture,Biology & Environmental Sciences
Life Sciences
Citazioni:
33
Recensione:
Indirizzi per estratti:
Indirizzo: Schuster, G Technion Israel Inst Technol, Dept Biol, IL-32000 Haifa, Israel Technion Israel Inst Technol Haifa Israel IL-32000 a, Israel
Citazione:
R. Rott et al., "Altering the 3 ' UTR endonucleolytic cleavage site of a Chlamydomonas chloroplast mRNA affects 3 '-end maturation in vitro but not in vivo", PLANT MOL B, 40(4), 1999, pp. 679-686

Abstract

The 3' ends of chloroplast mRNAs are produced by the processing of longer precursors. The 3' ends of most plastid mRNAs are located at, or several nucleotides downstream of, stem-loop structures, which act as 3'-end-processing signals and RNA stability elements. In chloroplasts of the green alga Chlamydomonas reinhardtii, 3'-end maturation of atpB mRNA involves endonucleolytic cleavage of the pre-mRNA at an AU-rich site located about 10 nucleotides downstream of the stem-loop structure. This cleavage is followed by exonucleolytic resection to generate the mature 3' end. In order to define critical nucleotides of the endonucleolytic cleavage site, we mutated its sequence. Incubation of synthetic atpB pre-RNAs containing these mutations in achloroplast protein extract resulted in the accumulation of 3'-end-processed products. However, in two cases where the AU-rich sequence of this site was replaced with a GC-rich one, the 3' end of the stable processing product differed from that of the wild-type product. To examine whether these mutations affected atpB mRNA processing or accumulation in vivo, the endogenous 3' UTR was replaced with mutated sequences by biolistic transformation ofChlamydomonas chloroplasts. Analysis of the resulting strains revealed that the accumulation of atpB mRNA was approximately equal to that of wild-type cells, and that a wild-type atpB 3' end was generated. These results imply that Chlamydomonas atpB 3' processing parallels the situation with other endonucleases such as Escherichia coli RNAse E, where specific sequences are required for correct in vitro processing, but in vivo these mutations canbe overcome.

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Documento generato il 25/01/20 alle ore 18:49:27