Reciprocal regulation of haem biosynthesis and the circadian clock in mammals

Abstract

The circadian clock is the central timing system that controls numerous physiological processes. In mammals, one such process is haem biosynthesis, which the clock controls through regulation of the rate-limiting enzyme aminolevulinate synthase 1 (Alas1)1,2. Several members of the core clock mechanism are PAS domain proteins, one of which, neuronal PAS 2 (NPAS2), has a haem-binding motif3,4. Indeed, haem controls activity of the BMAL1–NPAS2 transcription complex in vitro by inhibiting DNA binding in response to carbon monoxide3. Here we show that haem differentially modulates expression of the mammalian Period genes mPer1 and mPer2 in vivo by a mechanism involving NPAS2 and mPER2. Further experiments show that mPER2 positively stimulates activity of the BMAL1–NPAS2 transcription complex and, in turn, NPAS2 transcriptionally regulates Alas1. Vitamin B12 and haem compete for binding to NPAS2 and mPER2, but they have opposite effects on mPer2 and mPer1 expression in vivo. Our data show that the circadian clock and haem biosynthesis are reciprocally regulated and suggest that porphyrin-containing molecules are potential targets for therapy of circadian disorders.

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Figure 1: Haem modulates clock gene expression in liver.
Figure 2: NPAS2 is regulated by mPER2, and BMAL1–NPAS2 is a molecular target of mPER2.
Figure 3: NPAS2 is a principal regulator of ALAS1.
Figure 4: Haem and vitamin B12 binding by mPER2 and NPAS2.

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Acknowledgements

We thank S. L. McKnight for the Npas2m/m mice and NPAS2 antibody; M. Reick for technical comments; H. Okamura for the mPer1–luc and Clock expression constructs, P. Sassone-Corsi for the mPer2–luc expression plasmid; Z. Sun for suggesting the use of haem–agarose; and P. Hastings, R. Kellems and J. Lever for comments. This work was supported, in part, by a grant from the NIH (to C.C.L.).

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Correspondence to Cheng Chi Lee.

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The authors declare that they have no competing financial interests.

Supplementary information

Supplementary Figure 1

Synchronization of clock gene expression in NIH3T3 cells by haem. (PDF 1842 kb)

Supplementary Figure 2

a, Haem inhibits wheel-running activity during subjective night. b, Single-plotted wheel-running activity from mice injected at CT22. (PDF 4677 kb)

Supplementary Figure 3

NPAS2 and mPER2 mediate haem effects on mPer1 and mPer2 expression. (PDF 4813 kb)

Supplementary Figure 4

mPER1 is a minor regulator of NPAS2. (PDF 5068 kb)

Supplementary Figure 5

A model of reciprocal regulation between the circadian clock and haem biosynthesis. (PDF 4881 kb)

Supplementary Figure 6

c-Myc is regulated by NPAS2 and is modulated by haem. (PDF 3379 kb)

Legends for Supplementary Figures

c-Myc is regulated by NPAS2 and is modulated by haem. (DOC 25 kb)

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Kaasik, K., Chi Lee, C. Reciprocal regulation of haem biosynthesis and the circadian clock in mammals. Nature 430, 467–471 (2004). https://doi.org/10.1038/nature02724

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