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J Bacteriol, April 1998, p. 2167-2174, Vol. 180, No. 8
0021-9193/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.

A Period-Extender Gene, pex, That Extends the Period of the Circadian Clock in the Cyanobacterium Synechococcus sp. Strain PCC 7942

Shinsuke Kutsuna, Takao Kondo, Setsuyuki Aoki,dagger and Masahiro Ishiura*

Division of Biological Science, Graduate School of Science, Nagoya University, Chikusa-ku, Nagoya 464-01, and National Institute for Basic Biology, Okazaki, Aichi 444, Japan

Received 8 December 1997/Accepted 17 February 1998

We cloned the pS1K1 plasmid in the process of apparently "complementing" a circadian clock mutant of cyanobacterium Synechococcus sp. strain PCC 7942, SP22, which has a 22-h period (T. Kondo, N. F. Tsinoremas, S. S. Golden, C. H. Johnson, S. Kutsuna, and M. Ishiura, Science 266:1233-1236, 1994). Sequence analysis revealed that SP22 did not have a mutation in the genomic DNA segment carried on pS1K1, and the sp22 mutation was later found in a recently cloned new clock gene, kaiC. Therefore, the period-extender gene pex that was carried on pS1K1 was a suppressor gene for the sp22 mutation. The pex gene encoded a protein of 148 amino acid residues. No meaningful homologs were found in DNA or protein databases including the Synechocystis genome database. The pex gene was transcribed from 129 and 164 bp upstream of the translation initiation codon as 0.6-kb transcripts. The Pex protein was detected as a fusion protein with a molecular mass of 15 kDa by the epitope tag fusion method using a c-Myc epitope tag. Disruption of the pex gene in wild-type cells shortened the period of the rhythms by 1 h, although it did not affect other properties of the rhythms, whereas its overexpression extended the period by 3 h with a concomitant reduction in the amplitude of the rhythms. In various clock mutants examined, overexpression caused arrhythmicity. Thus, Pex is likely to function as a modifier of the circadian clock in Synechococcus.

* Corresponding author. Mailing address: Division of Biological Science, Graduate School of Science, Nagoya University, Furo-cho, Chikusa-ku, Nagoya 464-01, Japan. Phone: 81-52-789-2507. Fax: 81-52-789-2963. E-mail: ishiura{at}bio.nagoya-u.ac.jp.

dagger Present address: Division of Biological Informatics, Graduate School of Human Informatics, Nagoya University, Chikusa-ku, Nagoya 464-01, Japan.




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