Transcription of CABII is regulated by the biological clock in Chlamydomonas reinhardtii.
The small gene family encoding the chlorophyll a/b-binding proteins of photosystem II (CABII or lhcb) is known to exhibit circadian rhythms of mRNA abundance in Chlamydomonas reinhardtii. In this study we investigated the role of transcription in the phenomenon. We used as reporters Chlamydomonas genes that encode nitrate reductase (NITI) and arylsulfatase (ARS2) transcriptionally fused to sequences upstream of one of the CABII genes (called CABII-1). We found that both reporters exhibited the same circadian rhythm of mRNA abundance in phase, period, and amplitude as does the endogenous CABII-1 gene. We also evaluated the efficacy of arylsulfatase enzymatic activity as a reporter and found that its half-life is too long to make it a useful reporter of rhythmic transcription during a circadian or diurnal cycle. The amount of mRNA synthesis from the CABII-1 gene was examined by in vivo labeling experiments and a circadian rhythm in transcription rate was demonstrated. In vivo labeling also revealed a circadian rhythm of mRNA synthesis for the CABII gene family as a whole. The results from the transcriptional reporter assays together with the in vivo labeling experiments strongly support the conclusion that the biological clock regulates the transcriptional activity of the CABII-I gene, and moreover that regulation at the transcriptional level is the predominant mode by which the clock regulates this gene.
Department of Biology, Vanderbilt University, Nashville, TN/USA.
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S
Jacobshagen
K L
KL
Kindle
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CH
Johnson
D000818:Animals; D001192:Arylsulfatases / Q000096:biosynthesis / Q000235:genetics; D001683:Biological Clocks; D016825:Chlamydomonas reinhardtii / Q000235:genetics*; D002940:Circadian Rhythm; D056890:Eukaryota; D018628:Gene Dosage; D018506:Gene Expression Regulation, Plant; D017930:Genes, Reporter; D045342:Light-Harvesting Protein Complexes; D005810:Multigene Family; D050901:Nitrate Reductase; D009565:Nitrate Reductases / Q000096:biosynthesis / Q000235:genetics; D045322:Photosynthetic Reaction Center Complex Proteins / Q000096:biosynthesis / Q000235:genetics*; D045332:Photosystem II Protein Complex; D012333:RNA, Messenger / Q000032:analysis; D014158:Transcription, Genetic; D014170:Transformation, Genetic
D045342:Light-Harvesting Protein Complexes; D045322:Photosynthetic Reaction Center Complex Proteins; D045332:Photosystem II Protein Complex; D012333:RNA, Messenger; D009565:Nitrate Reductases; D050901:Nitrate Reductase; D001192:Arylsulfatases