title: Requirement for cytoplasmic protein synthesis during circadian peaks of transcription of chloroplast-encoded genes in Chlamydomonas.
abstract: Cycloheximide, an inhibitor of cytoplasmic translation, induced a rapid reduction of 70-80% in levels of mRNA for the chloroplast elongation factor Tu (tufA) in asynchronously growing Chlamydomonas. This effect was shown to be mainly transcriptional, and not restricted to tufA, as transcription of other chloroplast-encoded genes were cycloheximide-sensitive, although not all equally (psbA showed no more than 40% inhibition). Confirmatory evidence that the inhibition of chloroplast transcription was mainly due to blocking cytoplasmic translation was obtained with the cycloheximide-resistant mutant act1, and by using another translation inhibitor, anisomycin. In synchronously growing Chlamydomonas, chloroplast transcription is regulated by the circadian clock, with the daily peak occurring during the early light period. When cycloheximide was added during this period, transcription was inhibited, but not when it was added during the trough period (late light to early dark). Moreover, in synchronized cells switched to continuous light, the drug blocked the scheduled increase in tufA mRNA, but did not remove the pre-existing mRNA. These experiments define two functionally different types of chloroplast transcription in Chlamydomonas, basal (cycloheximide-insensitive) and clock-induced (cycloheximide-sensitive), and indicate that the relative contribution of each type to the overall transcription of a given gene are not identical for all genes. The results also provide evidence for nuclear regulation of chloroplast transcription, thereby obviating the need for an organellar clock, at least for these rhythms.
authors.affiliation: Section of Molecular Cell and Developmental Biology and Institute for Cellular and Molecular Biology, University of Texas at Austin, 78712, USA.
authors.forename: R
authors.initials: R
authors.lastname: Kawazoe
authors.forename: S
authors.initials: S
authors.lastname: Hwang
authors.forename: D L
authors.initials: DL
authors.lastname: Herrin
mesh_terms: D020418:Algal Proteins / Q000096:biosynthesis* / Q000235:genetics; D000818:Animals; D000841:Anisomycin / Q000494:pharmacology; D016825:Chlamydomonas reinhardtii / Q000187:drug effects / Q000235:genetics* / Q000378:metabolism; D002940:Circadian Rhythm; D003513:Cycloheximide / Q000494:pharmacology; D003593:Cytoplasm / Q000378:metabolism; D018742:DNA, Chloroplast / Q000235:genetics*; D004351:Drug Resistance; D005786:Gene Expression Regulation / Q000187:drug effects; D009154:Mutation; D010444:Peptide Elongation Factor Tu / Q000235:genetics; D014176:Protein Biosynthesis; D011500:Protein Synthesis Inhibitors / Q000494:pharmacology; D012333:RNA, Messenger / Q000187:drug effects / Q000235:genetics / Q000378:metabolism; D014158:Transcription, Genetic
chemical_list: D020418:Algal Proteins; D018742:DNA, Chloroplast; D011500:Protein Synthesis Inhibitors; D012333:RNA, Messenger; D000841:Anisomycin; D003513:Cycloheximide; D010444:Peptide Elongation Factor Tu