TY - JOUR
T1 - Light-induced activation of class II cyclobutane pyrimidine dimer photolyases
AU - Okafuji, Asako
AU - Biskup, Till
AU - Hitomi, Kenichi
AU - Getzoff, Elizabeth D.
AU - Kaiser, Gebhard
AU - Batschauer, Alfred
AU - Bacher, Adelbert
AU - Hidema, Jun
AU - Teranishi, Mika
AU - Yamamoto, Kazuo
AU - Schleicher, Erik
AU - Weber, Stefan
PY - 2010/5/4
Y1 - 2010/5/4
N2 - Light-induced activation of class II cyclobutane pyrimidine dimer (CPD) photolyases of Arabidopsis thaliana and Oryza sativa has been examined by UV/Vis and pulsed Davies-type electron-nuclear double resonance (ENDOR) spectroscopy, and the results compared with structure-known class I enzymes, CPD photolyase and (6-4) photolyase. By ENDOR spectroscopy, the local environment of the flavin adenine dinucleotide (FAD) cofactor is probed by virtue of proton hyperfine couplings that report on the electron-spin density at the positions of magnetic nuclei. Despite the amino-acid sequence dissimilarity as compared to class I enzymes, the results indicate similar binding motifs for FAD in the class II photolyases. Furthermore, the photoreduction kinetics starting from the FAD cofactor in the fully oxidized redox state, FADox, have been probed by UV/Vis spectroscopy. In Escherichia coli (class I) CPD photolyase, light-induced generation of FADH{radical dot} from FADox, and subsequently FADH- from FADH{radical dot}, proceeds in a step-wise fashion via a chain of tryptophan residues. These tryptophans are well conserved among the sequences and within all known structures of class I photolyases, but completely lacking from the equivalent positions of class II photolyase sequences. Nevertheless, class II photolyases show photoreduction kinetics similar to those of the class I enzymes. We propose that a different, but also effective, electron-transfer cascade is conserved among the class II photolyases. The existence of such electron transfer pathways is supported by the observation that the catalytically active fully reduced flavin state obtained by photoreduction is maintained even under oxidative conditions in all three classes of enzymes studied in this contribution.
AB - Light-induced activation of class II cyclobutane pyrimidine dimer (CPD) photolyases of Arabidopsis thaliana and Oryza sativa has been examined by UV/Vis and pulsed Davies-type electron-nuclear double resonance (ENDOR) spectroscopy, and the results compared with structure-known class I enzymes, CPD photolyase and (6-4) photolyase. By ENDOR spectroscopy, the local environment of the flavin adenine dinucleotide (FAD) cofactor is probed by virtue of proton hyperfine couplings that report on the electron-spin density at the positions of magnetic nuclei. Despite the amino-acid sequence dissimilarity as compared to class I enzymes, the results indicate similar binding motifs for FAD in the class II photolyases. Furthermore, the photoreduction kinetics starting from the FAD cofactor in the fully oxidized redox state, FADox, have been probed by UV/Vis spectroscopy. In Escherichia coli (class I) CPD photolyase, light-induced generation of FADH{radical dot} from FADox, and subsequently FADH- from FADH{radical dot}, proceeds in a step-wise fashion via a chain of tryptophan residues. These tryptophans are well conserved among the sequences and within all known structures of class I photolyases, but completely lacking from the equivalent positions of class II photolyase sequences. Nevertheless, class II photolyases show photoreduction kinetics similar to those of the class I enzymes. We propose that a different, but also effective, electron-transfer cascade is conserved among the class II photolyases. The existence of such electron transfer pathways is supported by the observation that the catalytically active fully reduced flavin state obtained by photoreduction is maintained even under oxidative conditions in all three classes of enzymes studied in this contribution.
KW - DNA repair
KW - Electron-nuclear double resonance
KW - Electron-paramagnetic resonance
KW - Flavoprotein
KW - Photolyase
KW - UV/blue light
UR - http://www.scopus.com/inward/record.url?scp=77952101898&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=77952101898&partnerID=8YFLogxK
U2 - 10.1016/j.dnarep.2010.01.014
DO - 10.1016/j.dnarep.2010.01.014
M3 - Article
C2 - 20227927
AN - SCOPUS:77952101898
VL - 9
SP - 495
EP - 505
JO - DNA Repair
JF - DNA Repair
SN - 1568-7864
IS - 5
ER -