Circadian Output, Input, and Intracellular Oscillators: Insights into the Circadian Systems of Single Cells

  1. J. J. Loros*†,
  2. J. C. Dunlap,
  3. L. F. Larrondo,
  4. M. Shi,
  5. W. J. Belden,
  6. V. D. Gooch,
  7. C.-H. Chen,
  8. C. L. Baker,
  9. A. Mehra,
  10. H. V. Colot,
  11. C. Schwerdtfeger,
  12. R. Lambreghts,
  13. P. D. Collopy,
  14. J. J. Gamsby, and
  15. C. I. Hong
  1. *Department of Biochemistry, Dartmouth Medical School, Hanover, New Hampshire 03755
  2. Department of Genetics, Dartmouth Medical School, Hanover, New Hampshire 03755
  3. Division of Science and Mathematics, University of Minnesota, Morris, Minnesota 56267

Abstract

Circadian output comprises the business end of circadian systems in terms of adaptive significance. Work on Neurospora pioneered the molecular analysis of circadian output mechanisms, and insights from this model system continue to illuminate the pathways through which clocks control metabolism and overt rhythms. In Neurospora, virtually every strain examined in the context of rhythms bears the band allele that helps to clarify the overt rhythm in asexual development. Recent cloning of band showed it to be an allele of ras-1 and to affect a wide variety of signaling pathways yielding enhanced light responses and asexual development. These can be largely phenocopied by treatments that increase levels of intracellular reactive oxygen species. Although output is often unidirectional, analysis of the prd-4 gene provided an alternative paradigm in which output feeds back to affect input. prd-4 is an allele of checkpoint kinase-2 that bypasses the requirement for DNA damage to activate this kinase; FRQ is normally a substrate of activated Chk2, so in Chk2PRD-4, FRQ is precociously phosphorylated and the clock cycles more quickly. Finally, recent adaptation of luciferase to fully function in Neurospora now allows the core FRQ/WCC feedback loop to be followed in real time under conditions where it no longer controls the overt rhythm in development. This ability can be used to describe the hierarchical relationships among FRQ-Less Oscillators (FLOs) and to see which are connected to the circadian system. The nitrate reductase oscillator appears to be connected, but the oscillator controlling the long-period rhythm elicited upon choline starvation appears completely disconnected from the circadian system; it can be seen to run with a very long noncompensated 60–120-hour period length under conditions where the circadian FRQ/WCC oscillator continues to cycle with a fully compensated circadian 22-hour period.

Footnotes

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