Photolyase is a photoenzyme that uses the energy of blue light to reverse UV-induced DNA damage in many organisms. Cryptochrome is a recently discovered blue-light photoreceptor that regulates the circadian clock in animals (and plants) and growth and development in plants. Both proteins have similar structural architectures but with totally different functions. We have recently elucidated the repair mechanisms and photocycles of cyclobutane pyrimidine dimer by a class of microbial photolyases (class-I) and of pyrimidine-pyrimidone (6-4) photoproduct by (6-4) photolyases. Several new classes of photolyases have been recently discovered with novel active sites and some new functions have also been lately observed. Thus, the first objective (Aim 1) of the project is to systematically characterize the repair of cyclobutane pyrimidine dimer by all other three classes of photolyases and the new function of Dewar repair by (6-4) photolyases. Such systematic investigations will obtain the molecular understanding of detrimental effects of UV radiation on the biosphere. Cryptochrome has been heavily studied by molecular genetics but the mechanistic investigation and understanding are simply lacking. Even the active (functional) redox state of cofactor flavin in cryptochrome is unknown yet. It is only known that cryptochrome proceeds to conformational changes to trigger downstream signal transduction upon blue-light photoreception. Thus, the second objective (Aim 2) is to systematically investigate the redox state(s) and related photochemistry, determine the active state in vivo in plant and animal cryptochromes, and characterize subsequent conformation dynamics and related interactions with downstream proteins. These investigations will uncover the primary process of initial signal transduction and reveal the reaction mechanism and photocycle of cryptochrome. To achieve these goals, we integrate state-of-the-art laser spectroscopy and biochemistry/molecular biology and follow the entire functional evolution of DNA repair and initial signaling of the complex processes with femtosecond temporal resolution and single-residue spatial resolution. The new knowledge obtained from this work on photolyase and cryptochrome is significant to the DNA repair and biological clock fields and, more importantly, is critical to practical applications of rug design for a series of diseases such as skin cancer and mental disorder.

Public Health Relevance

Blue light is used to repair DNA damage caused by UV irradiation through a photoenzyme (photolyase) and to synchronize the circadian clock in animals or to regulate growth and development in plants via signal transduction by a photoreceptor (cryptochrome). DNA damage could lead to skin cancer and circadian rhythms are related to many mental diseases. Here, we develop a novel method by integrating femtosecond laser spectroscopy and biochemistry/molecular biology to systematically characterize the repair dynamics of damaged DNA by photolyase and the primary signaling process by cryptochrome. The new knowledge from these studies is fundamental to the DNA-repair and signal transduction fields and also significant to a series of potential applications suh as drug design and prevention of skin cancer and mental disorder.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
2R01GM074813-06
Application #
8438981
Study Section
Macromolecular Structure and Function B Study Section (MSFB)
Program Officer
Smith, Ward
Project Start
2005-06-01
Project End
2017-04-30
Budget Start
2013-08-01
Budget End
2014-04-30
Support Year
6
Fiscal Year
2013
Total Cost
$290,072
Indirect Cost
$98,266
Name
Ohio State University
Department
Chemistry
Type
Schools of Arts and Sciences
DUNS #
832127323
City
Columbus
State
OH
Country
United States
Zip Code
43210
Tan, Chuang; Guo, Lijun; Ai, Yuejie et al. (2014) Direct determination of resonance energy transfer in photolyase: structural alignment for the functional state. J Phys Chem A 118:10522-30
Liu, Zheyun; Zhang, Meng; Guo, Xunmin et al. (2013) Dynamic determination of the functional state in photolyase and the implication for cryptochrome. Proc Natl Acad Sci U S A 110:12972-7
Liu, Zheyun; Tan, Chuang; Guo, Xunmin et al. (2013) Determining complete electron flow in the cofactor photoreduction of oxidized photolyase. Proc Natl Acad Sci U S A 110:12966-71
Liu, Zheyun; Guo, Xunmin; Tan, Chuang et al. (2012) Electron tunneling pathways and role of adenine in repair of cyclobutane pyrimidine dimer by DNA photolyase. J Am Chem Soc 134:8104-14
Kao, Ya-Ting; Song, Qin-Hua; Saxena, Chaitanya et al. (2012) Dynamics and mechanism of DNA repair in a biomimetic system: flavin-thymine dimer adduct. J Am Chem Soc 134:1501-3
Liu, Zheyun; Tan, Chuang; Guo, Xunmin et al. (2011) Dynamics and mechanism of cyclobutane pyrimidine dimer repair by DNA photolyase. Proc Natl Acad Sci U S A 108:14831-6
Hassanali, Ali A; Zhong, Dongping; Singer, Sherwin J (2011) An AIMD study of the CPD repair mechanism in water: reaction free energy surface and mechanistic implications. J Phys Chem B 115:3848-59
Li, Xu; Wang, Qin; Yu, Xuhong et al. (2011) Arabidopsis cryptochrome 2 (CRY2) functions by the photoactivation mechanism distinct from the tryptophan (trp) triad-dependent photoreduction. Proc Natl Acad Sci U S A 108:20844-9
Hassanali, Ali A; Zhong, Dongping; Singer, Sherwin J (2011) An AIMD study of CPD repair mechanism in water: role of solvent in ring splitting. J Phys Chem B 115:3860-71
Li, Jiang; Liu, Zheyun; Tan, Chuang et al. (2010) Dynamics and mechanism of repair of ultraviolet-induced (6-4) photoproduct by photolyase. Nature 466:887-890

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