Organisms have developed complex regulatory systems in order to couple metabolism and cell development with changes in their environment. Such adaptation is well demonstrated by circadian clocks, which synchronize cellular responses to the diurnal light cycle. Due to their central role in pacing metabolism, biological clocks impact many aspects of human physiology, behavior and the treatment of disease. Although genetics and cell biology have made great progress in understanding the function of clock genes, the molecular mechanisms that compose the underlying transcriptional feedback loops, and their light entrapment, remain largely unknown. Primary photoreceptors and their targets have been identified in eukaryotic model systems such as Neurospora, (filamentous fungi) and Drosophila (flies). Preliminary characterization of light-sensors white collar 1 (WC-1) and vivid (WD) from Neurospora and cryptochrome (CRY) from Drosophila, has defined structures and reactivities that implicate both flavin photochemistry and thiolate redox chemistry in the activation of Per-Arnt-Sim (PAS) domain-containing transcription factors. Crystallographic and spectroscopic characterization of how large-scale protein conformational changes propagate from cofactor photochemistry to generate complex alterations in protein/protein interactions will provide a molecular understanding for clock function. Studies of circadian clock proteins have been hampered by the complexity of the cellular responses, the presence of multiple entrainment mechanisms, and the lack of structures for key proteins. High-resolution structures of these photoreceptors in various states of activation and in complex with their downstream partners will delineate individual residues important for cellular reactivity. Both in vivo and in vitro analysis of variant clock proteins designed to decouple their properties will help fragment entrainment pathways and identify additional levels of clock control, such as response to cellular redox potentials. Parallel investigations of related mammalian clock proteins will reveal mechanisms of action that will aid in the prediction and perturbation of function. Ultimately, such studies of clock proteins will provide a molecular description of behavior that promises advancements in the treatment of mental disorders and many other maladies.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
5R01GM079679-04
Application #
7849652
Study Section
Macromolecular Structure and Function B Study Section (MSFB)
Program Officer
Wehrle, Janna P
Project Start
2007-06-01
Project End
2011-08-31
Budget Start
2010-06-01
Budget End
2011-08-31
Support Year
4
Fiscal Year
2010
Total Cost
$280,315
Indirect Cost
Name
Cornell University
Department
Chemistry
Type
Schools of Arts and Sciences
DUNS #
872612445
City
Ithaca
State
NY
Country
United States
Zip Code
14850
Conrad, Karen S; Hurley, Jennifer M; Widom, Joanne et al. (2016) Structure of the frequency-interacting RNA helicase: a protein interaction hub for the circadian clock. EMBO J 35:1707-19
Ganguly, Abir; Manahan, Craig C; Top, Deniz et al. (2016) Changes in active site histidine hydrogen bonding trigger cryptochrome activation. Proc Natl Acad Sci U S A 113:10073-8
Yee, Estella F; Diensthuber, Ralph P; Vaidya, Anand T et al. (2015) Signal transduction in light-oxygen-voltage receptors lacking the adduct-forming cysteine residue. Nat Commun 6:10079
Merz, Gregory E; Borbat, Peter P; Pratt, Ashley J et al. (2014) Copper-based pulsed dipolar ESR spectroscopy as a probe of protein conformation linked to disease states. Biophys J 107:1669-74
Crane, Brian R; Young, Michael W (2014) Interactive features of proteins composing eukaryotic circadian clocks. Annu Rev Biochem 83:191-219
Conrad, Karen S; Manahan, Craig C; Crane, Brian R (2014) Photochemistry of flavoprotein light sensors. Nat Chem Biol 10:801-9
Conrad, Karen S; Bilwes, Alexandrine M; Crane, Brian R (2013) Light-induced subunit dissociation by a light-oxygen-voltage domain photoreceptor from Rhodobacter sphaeroides. Biochemistry 52:378-91
Vaidya, Anand T; Top, Deniz; Manahan, Craig C et al. (2013) Flavin reduction activates Drosophila cryptochrome. Proc Natl Acad Sci U S A 110:20455-60
Levy, Colin; Zoltowski, Brian D; Jones, Alex R et al. (2013) Updated structure of Drosophila cryptochrome. Nature 495:E3-4
Vaidya, Anand T; Chen, Chen-Hui; Dunlap, Jay C et al. (2011) Structure of a light-activated LOV protein dimer that regulates transcription. Sci Signal 4:ra50

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