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A systematic forward genetic analysis identified components of the Chlamydomonas circadian system

¹ Center for Gene Research, Nagoya University, Furo, Chikusa, Nagoya 464-8602, Japan; ² Division of Biological Science, Graduate School of Science, Nagoya University, Furo, Chikusa, Nagoya 464-8602, Japan; ³ Laboratory of Chemistry, School of Medicine, Teikyo University, Ohtsuka, Hachioji, Tokyo 192-0395, Japan

The molecular bases of circadian clocks have been studied in animals, fungi, bacteria, and plants, but not in eukaryotic algae. To establish a new model for molecular analysis of the circadian clock, here we identified a large number of components of the circadian system in the eukaryotic unicellular alga Chlamydomonas reinhardtii by a systematic forward genetic approach. We isolated 105 insertional mutants that exhibited defects in period, phase angle, and/or amplitude of circadian rhythms in bioluminescence derived from a luciferase reporter gene in their chloroplast genome. Simultaneous measurement of circadian rhythms in bioluminescence and growth rate revealed that some of these mutants had defects in the circadian clock itself, whereas one mutant had a defect in a specific process for the chloroplast bioluminescence rhythm. We identified 30 genes (or gene loci) that would be responsible for rhythm defects in 37 mutants. Classification of these genes revealed that various biological processes are involved in regulation of the chloroplast rhythmicity. Amino acid sequences of six genes that would have crucial roles in the circadian clock revealed features of the Chlamydomonas clock that have both partially plant-like and original components.

[Keywords: Chlamydomonas reinhardtii; circadian rhythm; bioluminescence; insertional mutagenesis; clock gene]]

Received January 11, 2008; revised version accepted January 28, 2008.

E-MAIL ishiura{at}gene.nagoya-u.ac.jp; FAX 81-52-789-4526.

Supplemental material is available at http://www.genesdev.org.

Article published online ahead of print. Article and publication date are online at http://www.genesdev.org/cgi/doi/10.1101/gad.1650408.

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