Regulation and molecular structure of a circadian oscillating protein located in the cell membrane of the prokaryote Synechococcus RF-1.

Abstract

When a light/light-adapted culture of Synechococcus RF-1 is exposed to a diurnal light/dark regimen, the synthesis of more than ten of its polypeptides is known to become entrained to a circadian oscillating pattern which persists for some time under free-running conditions. One of the circadian oscillating polypeptides, COP23, was found to be located in the cell membrane. The rate of COP23 synthesis is controlled at the transcription level. In addition to the protein synthesis rate, the content of COP23 also exhibited a circadian rhythm. Pulse labeling with [35S]methionine revealed that COP23 was relatively stable in an arrhythmic culture. However, the exposure of Synechococcus RF-1 to a light/dark regimen induced not only a circadian synthesis rhythm, but also a rapid degradation of COP23 protein at a defined period of time. The induction of rapid protein degradation was prevented by the presence of chloramphenicol. The gene encoding the COP23 polypeptide has been cloned and sequenced. The amino acid sequence derived from the open-reading frame revealed that a signal peptide (28 amino acids) does not appear to be part of the mature COP23. The mature COP23 does not have a membrane-associated segment, and it is suggested to be a peripheral molecule. With respect to their DNA base sequence and protein amino acid sequence, none of the proteins documented in the EMBL and PC/Gene data bases are significantly homologous with the COP23 molecule.