TY - JOUR
T1 - Novel DNA binding protiens highly specific to UV-damaged DNA sequences from embryos of Drosophila melanogaster
AU - Kai, Mihoko
AU - Takahashi, Toshiaki
AU - Todo, Takeshi
AU - Sakaguchi, Kengo
N1 - Copyright:
Copyright 2010 Elsevier B.V., All rights reserved.
PY - 1995/7/25
Y1 - 1995/7/25
N2 - Three new proteins which selectively bind to UVdamaged DNA were identified and purified to near homogeneity from UV-lrradiated Drosophila melanogaster embryos through several column chromatographles. These proteins, tentatively designated as D-DDB P1, P2 and P3, can be identified as different complex bands in a gel shift assay by using UVirradiated TC-31 probe DNA. Analysis of the purified D-DDB P1 fraction by native or SDS-polyacrylamlde gel electrophoresis and FPLC-Superose 6 gel filtration demonstrated that it is a monomer protein which Is a 30 kDa polypeptlde. The D-DDB P2 protein is a monopolypeptide with a molecular mass of 14 kDa. Both D-DDB P1 and P2 highly prefer binding to UV-irradiated DNA, and have almost no affinity for non-irradiated DNA. Gel shift assays with either UV-irradiated DNA probes demonstrated that D-DDB P1 may show a preference for binding to (6-4) photoproducts, while D-DDB P2 may prefer binding to pyrimidine dimers. Both these proteins require magnesium ions for binding. D-DDB P1 is an ATP-preferent protein. These findings are discussed in relation to two recently described [Todo and Ryo (1991) Mutat. Res., 273, 85-93; Todo et al. (1993) Nature, 361, 371-374] DNA-binding factors from Drosophila cell extracts. A possible role for these DNA-binding proteins in lesion recognition and DNA repair of UV-induced photoproducts is discussed.
AB - Three new proteins which selectively bind to UVdamaged DNA were identified and purified to near homogeneity from UV-lrradiated Drosophila melanogaster embryos through several column chromatographles. These proteins, tentatively designated as D-DDB P1, P2 and P3, can be identified as different complex bands in a gel shift assay by using UVirradiated TC-31 probe DNA. Analysis of the purified D-DDB P1 fraction by native or SDS-polyacrylamlde gel electrophoresis and FPLC-Superose 6 gel filtration demonstrated that it is a monomer protein which Is a 30 kDa polypeptlde. The D-DDB P2 protein is a monopolypeptide with a molecular mass of 14 kDa. Both D-DDB P1 and P2 highly prefer binding to UV-irradiated DNA, and have almost no affinity for non-irradiated DNA. Gel shift assays with either UV-irradiated DNA probes demonstrated that D-DDB P1 may show a preference for binding to (6-4) photoproducts, while D-DDB P2 may prefer binding to pyrimidine dimers. Both these proteins require magnesium ions for binding. D-DDB P1 is an ATP-preferent protein. These findings are discussed in relation to two recently described [Todo and Ryo (1991) Mutat. Res., 273, 85-93; Todo et al. (1993) Nature, 361, 371-374] DNA-binding factors from Drosophila cell extracts. A possible role for these DNA-binding proteins in lesion recognition and DNA repair of UV-induced photoproducts is discussed.
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U2 - 10.1093/nar/23.14.2600
DO - 10.1093/nar/23.14.2600
M3 - Article
C2 - 7651820
AN - SCOPUS:0029157371
SN - 0305-1048
VL - 23
SP - 2600
EP - 2607
JO - Nucleic acids research
JF - Nucleic acids research
IS - 14
ER -