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A chloroplast transcript lacking the 3' inverted repeat is degraded by 3'-->5' exoribonuclease activity.

RNA (New York, N.Y.)

genetics, metabolism, Animals, Base Sequence, Chlamydomonas reinhardtii, growth & development, physiology, Chloroplasts, Escherichia coli, enzymology, Exoribonucleases, Transformation, Genetic, Guanosine, chemistry, Models, Genetic, Molecular Sequence Data, Photosynthesis, Plasmids, Polydeoxyribonucleotides, RNA Processing, Post-Transcriptional, Repetitive Sequences, Nucleic Acid, Transcription, Genetic, Adenosine Triphosphatases

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      Chlamydomonas reinhardtii strains harboring deletions of the chloroplast atpB 3' inverted repeat (IR) are weakly phototrophic due to reduced accumulation of discrete atpB transcripts and the chloroplast ATPase beta-subunit protein. A sequence of 18 guanosine residues, which can impede a 3'-->5' exoribonuclease in vitro, is able to substitute for the atpB IR in vivo. Strains containing the poly-guanosine tract in place of the atpB 3' IR are phototrophic and accumulate near wild-type levels of discrete atpB transcripts and the ATPase beta-subunit protein. Because these atpB transcripts contain the 18 guanosine residues, and the poly-guanosine tract is not a terminator of transcription, the accumulation of discrete atpB transcripts is likely the result of impediment of 3'-->5' exoribonuclease activity. These findings support a model in which atpB transcripts lacking the 3' IR are degraded by 3'-->5' exoribonuclease activity, and demonstrate that the poly-guanosine tract can be used to study chloroplast RNA metabolism in vivo.

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