Genetic and phylogenetic characterization of the type II cyclobutane pyrimidine dirner photolyases encoded by Leporipoxviruses

被引:22
作者
Bennett, CJ [1 ]
Webb, M [1 ]
Willer, DO [1 ]
Evans, DH [1 ]
机构
[1] Univ Guelph, Dept Mol Biol & Genet, Guelph, ON N1G 2W1, Canada
基金
加拿大健康研究院; 加拿大自然科学与工程研究理事会;
关键词
cyclobutane pyrimidine dimer; DNA repair; CPD photolyase; Shope fibroma virus; UV light; myxoma virus; virus phylogeny;
D O I
10.1016/S0042-6822(03)00512-9
中图分类号
Q93 [微生物学];
学科分类号
071005 ; 100705 ;
摘要
Shope fibroma virus and myxoma virus encode proteins predicted to be Type II photolyases. These are enzymes that catalyze light-dependent repair of cyclobutane pyrimidine dimers (CPDs). When the Shope fibroma virus S127L gene was expressed in an Escherichia coli strain lacking functional CPD repair pathways, the expressed gene protected the bacteria from 70-75% of the ultraviolet (UV) light-induced cytotoxic DNA damage. This proportion suggests that Leporipoxvirus photolyases can only repair CPDs, which typically comprise similar to70% of the damage caused by short wavelength UV light. To test whether these enzymes can protect virus genomes from UV, we exposed virus suspensions to UV-C light followed by graded exposure to filtered visible light. Viruses encoding a deletion of the putative photolyase gene were unable to photoreactivate UV damage while this treatment again eliminated 70-90% of the lethal photoproducts in wild-type viruses. Western blotting detected photolyase protein in extracts prepared from purified virions and it can be deduced that the poxvirion interior must be fluid enough to permit diffusion of this similar to50-kDa DNA-binding protein to the sites where it catalyzes photoreactivation. Photolyase promoters are difficult to categorize using bioinformatics methods, as they do not obviously resemble any of the known poxvirus promoter motifs. By fusing the SFV promoter to DNA encoding a luciferase open reading frame, the photolyase promoter was found to exhibit very weak late promoter activity. These data show that the genomes of Leporipox viruses, similar to that of fowlpox virus, encode catalytically active photolyases. Phylogenetic studies also confirmed the monophyletic origin of poxviruses and suggest an ancient origin for these genes and perhaps poxviruses. (C) 2003 Elsevier Inc. All rights reserved.
引用
收藏
页码:10 / 19
页数:10
相关论文
共 40 条
[1]   The genome of fowlpox virus [J].
Afonso, CL ;
Tulman, ER ;
Lu, Z ;
Zsak, L ;
Kutish, GF ;
Rock, DL .
JOURNAL OF VIROLOGY, 2000, 74 (08) :3815-3831
[2]   The genome of Melanoplus sanguinipes Entomopoxvirus [J].
Afonso, CL ;
Tulman, ER ;
Lu, Z ;
Oma, E ;
Kutish, GF ;
Rock, DL .
JOURNAL OF VIROLOGY, 1999, 73 (01) :533-552
[3]   Complete genomic sequence of the Amsacta moorei entomopoxvirus:: Analysis and comparison with other poxviruses [J].
Bawden, AL ;
Glassberg, KJ ;
Diggans, J ;
Shaw, R ;
Farmerie, W ;
Moyer, RW .
VIROLOGY, 2000, 274 (01) :120-139
[4]  
BRASH DE, 1985, J BIOL CHEM, V260, P1438
[5]   The complete DNA sequence of myxoma virus [J].
Cameron, C ;
Hota-Mitchell, S ;
Chen, L ;
Barrett, J ;
Cao, JX ;
Macaulay, C ;
Willer, D ;
Evans, D ;
McFadden, G .
VIROLOGY, 1999, 264 (02) :298-318
[6]   Leporipoxvirus Cu-Zn superoxide dismutase homologs inhibit cellular superoxide dismutase, but are not essential for virus replication or virulence [J].
Cao, JX ;
Teoh, MLT ;
Moon, M ;
McFadden, G ;
Evans, DH .
VIROLOGY, 2002, 296 (01) :125-135
[7]   In vivo formation and repair of cyclobutane pyrimidine dimers and 6-4 photoproducts measured at the gene and nucleotide level in Escherichia coli [J].
Chandrasekhar, D ;
Van Houten, B .
MUTATION RESEARCH-FUNDAMENTAL AND MOLECULAR MECHANISMS OF MUTAGENESIS, 2000, 450 (1-2) :19-40
[8]   TUMORIGENIC POXVIRUSES - CONSTRUCTION OF THE COMPOSITE PHYSICAL MAP OF THE SHOPE FIBROMA VIRUS GENOME [J].
DELANGE, AM ;
MACAULAY, C ;
BLOCK, W ;
MUELLER, T ;
MCFADDEN, G .
JOURNAL OF VIROLOGY, 1984, 50 (02) :408-416
[9]   Determining divergence times of the major kingdoms of living organisms with a protein clock [J].
Doolittle, RF ;
Feng, DF ;
Tsang, S ;
Cho, G ;
Little, E .
SCIENCE, 1996, 271 (5248) :470-477
[10]   Individual determination of the yield of the main UV-induced dimeric pyrimidine photoproducts in DNA suggests a high mutagenicity of CC photolesions [J].
Douki, T ;
Cadet, J .
BIOCHEMISTRY, 2001, 40 (08) :2495-2501