DNA-Binding Properties of the Antibody Specific for the Dewar Photoproduct of Thymidylyl-(3′′-5′′)-Thymidine

A monoclonal antibody (DEM-1) specific for the Dewar photoproduct is used for detection and quantification of photolesions in DNA. To help understand the molecular recognition of damaged DNA by the antibody protein, we have cloned and sequenced the variable region genes of DEM-1. We have also prepared Fab fragments of DEM-1 (DEM1Fab), and synthesized two kinds of 3′-biotinylated oligonucleotides of different lengths containing a central Dewar photoproduct of TpT to analyze the effects of the antigen size on the binding rates by means of surface plasmon resonance (SPR). Results obtained from SPR analyses suggest that DEM1Fab may recognize tetranucleotide unit as the epitope.

[1]  Y. Komatsu,et al.  Antigen structural requirements for recognition by a cyclobutane thymine dimer-specific monoclonal antibody. , 1997, Nucleic acids research.

[2]  O. Nikaido,et al.  ESTABLISHMENT and CHARACTERIZATION OF A MONOCLONAL ANTIBODY RECOGNIZING THE DEWAR ISOMERS OF(6–4)PHOTOPRODUCTS , 1993, Photochemistry and photobiology.

[3]  E. Ohtsuka,et al.  Synthesis of a Phosphoramidite Coupling Unit of the Pyrimidine (6−4) Pyrimidone Photoproduct and Its Incorporation into Oligodeoxynucleotides , 1996 .

[4]  D R Burton,et al.  Generation of a large combinatorial library of the immunoglobulin repertoire in phage lambda. , 1989, Science.

[5]  R. Wood.  DNA repair in eukaryotes. , 1996, Annual review of biochemistry.

[6]  J. E. Leclerc,et al.  The thymine-thymine pyrimidine-pyrimidone(6-4) ultraviolet light photoproduct is highly mutagenic and specifically induces 3' thymine-to-cytosine transitions in Escherichia coli. , 1991, Proceedings of the National Academy of Sciences of the United States of America.

[7]  Y. Komatsu,et al.  Crystal structure of the 64M-2 antibody Fab fragment in complex with a DNA dT(6-4)T photoproduct formed by ultraviolet radiation. , 2000, Journal of molecular biology.

[8]  E. Ohtsuka,et al.  Tryptophan H33 plays an important role in pyrimidine (6-4) pyrimidone photoproduct binding by a high-affinity antibody. , 1999, Protein engineering.

[9]  Y. Komatsu,et al.  31P NMR study of the interactions between oligodeoxynucleotides containing (6‐4) photoproduct and Fab fragments of monoclonal antibodies specific for (6‐4) photoproduct , 1998, FEBS letters.

[10]  T. Torizawa,et al.  Specificities and rates of binding of anti-(6-4) photoproduct antibody fragments to synthetic thymine photoproducts. , 1998, Journal of biochemistry.

[11]  O. Nikaido,et al.  Establishment of a monoclonal antibody recognizing cyclobutane-type thymine dimers in DNA: a comparative study with 64M-1 antibody specific for (6-4)photoproducts. , 1991, Mutation research.

[12]  R. Baan,et al.  DETECTION OF CYCLOBUTANE THYMINE DIMERS IN DNA OF HUMAN CELLS WITH MONOCLONAL ANTIBODIES RAISED AGAINST A THYMINE DIMER‐ CONTAINING TETRANUCLEOTIDE , 1988, Photochemistry and photobiology.

[13]  Y. Komatsu,et al.  Conformational multiplicity of the antibody combining site of a monoclonal antibody specific for a (6-4) photoproduct. , 1999, Journal of molecular biology.

[14]  JohnB . Taylor,et al.  DNA, light, and Dewar pyrimidinones: the structure and biological significance to TpT3 , 1987 .

[15]  I. Shimada,et al.  Proteolytic fragmentation with high specificity of mouse immunoglobulin G. Mapping of proteolytic cleavage sites in the hinge region. , 1995, Journal of immunological methods.

[16]  T. Toyo’oka,et al.  Isolation and characterization of cysteine-containing regions of proteins using 4-(aminosulfonyl)-7-fluoro-2,1,3-benzoxadiazole and high-performance liquid chromatography. , 1985, Analytical chemistry.

[17]  C A Smith,et al.  Preparation and characterization of a set of deoxyoligonucleotide 49-mers containing site-specific cis-syn, trans-syn-I, (6-4), and Dewar photoproducts of thymidylyl(3'-->5')-thymidine. , 1993, The Journal of biological chemistry.

[18]  H. Krokan,et al.  U.v.-induced DNA damage and its repair in human skin in vivo studied by sensitive immunohistochemical methods. , 1983, Carcinogenesis.

[19]  W. Baeyens,et al.  Availability of fluorogenic reagents having a benzofurazan structure in the biosciences , 1994 .

[20]  O. Nikaido,et al.  Antibodies specific for (6-4) DNA photoproducts: cloning, antibody modeling and construction of a single-chain Fv derivative. , 1998, Biochimica et biophysica acta.

[21]  A. Sancar.  DNA excision repair. , 1996, Annual review of biochemistry.

[22]  O. Nikaido,et al.  SIMULTANEOUS ESTABLISHMENT OF MONOCLONAL ANTIBODIES SPECIFIC FOR EITHER CYCLOBUTANE PYRIMIDINE DIMER OR (6‐4)PHOTOPRODUCT FROM THE SAME MOUSE IMMUNIZED WITH ULTRAVIOLET‐IRRADIATED DNA , 1991, Photochemistry and photobiology.

[23]  D. Mitchell,et al.  The development of a radioimmunoassay for the detection of photoproducts in mammalian cell DNA. , 1981, Biochimica et biophysica acta.

[24]  E. Friedberg,et al.  DNA Repair and Mutagenesis , 2006 .

[25]  Y. Komatsu,et al.  DNA binding mode of the Fab fragment of a monoclonal antibody specific for cyclobutane pyrimidine dimer. , 2000, Nucleic acids research.

[26]  A. Wani,et al.  ANTIBODIES TO UV IRRADIATED DNA: THE MONITORING OF DNA DAMAGE BY ELISA AND INDIRECT IMMUNOFLUORESCENCE , 1984, Photochemistry and photobiology.

[27]  E. Kabat,et al.  Sequences of proteins of immunological interest , 1991 .

[28]  Y. Komatsu,et al.  Efficient chemical synthesis of a pyrimidine (6–4) pyrimidone photoproduct analog and its properties , 2000 .

[29]  M. Taussig,et al.  Characterization of germ-line genes of the VGAM3.8 VH gene family from BALB/c mice. , 1992, Journal of immunology.

[30]  D. Pisetsky,et al.  Anti-DNA antibodies from autoimmune mice arise by clonal expansion and somatic mutation , 1990, The Journal of experimental medicine.

[31]  O. Nikaido,et al.  The role of surface lysines in pyrimidine (6-4) pyrimidone photoproduct binding by a high-affinity antibody. , 1998, Protein engineering.

[32]  O. Nikaido,et al.  Establishment of a monoclonal antibody recognizing ultraviolet light-induced (6-4) photoproducts. , 1988, Mutation research.

[33]  R. Setlow.  Repair deficient human disorders and cancer , 1978, Nature.

[34]  J. Sambrook,et al.  Molecular Cloning: A Laboratory Manual , 2001 .

[35]  C. Smith,et al.  Mutation spectra of M13 vectors containing site-specific Cis-Syn, Trans-Syn-I, (6-4), and Dewar pyrimidone photoproducts of thymidylyl-(3'-->5')-thymidine in Escherichia coli under SOS conditions. , 1996, Biochemistry.

[36]  W. Haseltine,et al.  Alkaline lability of fluorescent photoproducts produced in ultraviolet light-irradiated DNA. , 1982, The Journal of biological chemistry.