Control of mRNA stability in chloroplasts by 3′ inverted repeats: effects of stem and loop mutations on degradation ofpsbA mRNAin vitro
| Autor: | Cynthia C. Adams, David B. Stern |
|---|---|
| Rok vydání: | 1990 |
| Předmět: |
Base Composition
Messenger RNA RNA Stability Chloroplasts Base Sequence Transcription Genetic biology Inverted repeat Molecular Sequence Data Mutant RNA Templates Genetic Plants Molecular biology Endonuclease Gene expression Mutagenesis Site-Directed Genetics biology.protein Nucleic Acid Conformation RNA Messenger Cloning Molecular Gene Software Repetitive Sequences Nucleic Acid |
| Zdroj: | Nucleic Acids Research. 18:6003-6010 |
| ISSN: | 1362-4962 0305-1048 |
| DOI: | 10.1093/nar/18.20.6003 |
| Popis: | To investigate the role of mRNA 3' inverted repeats (IRs) in stabilizing plant chloroplast mRNAs, we have measured the processing and stability of wild-type and mutant RNAs corresponding to the 3' end of the spinach chloroplast psbA mRNA. wild-type and mutant 3' IR-RNA precursors were processed at similar rates in a homologous in vitro system, but RNAs with either a mutant loop sequence CUUCGG or a specific base substitution in the IR exhibited an enhanced accumulation of mature product. Incubation of mature products in the in vitro system demonstrated that this was due to an increased stability of the product. These mutant RNAs displayed the same order of stabilities when their decay was measured following electroporation into intact chloroplasts. We found that the in vitro system contains an endonuclease activity that cleaves the wild-type 3' IR-RNA within the loop and also in single-stranded regions, suggesting a possible role for the loop sequence in determining RNA longevity in vitro. Interestingly, the altered loop sequence CUUCGG, which enhances RNA stability in bacteria (1), prolonged the half-life of psbA 3' IR-RNA in vitro and also resulted in an altered endonuclease cleavage pattern. Such nucleases could potentially play an important role in plastid mRNA decay in vivo. |
| Databáze: | OpenAIRE |
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