Partial complementation of the UV sensitivity of E. coli and yeast excision repair mutants by the cloned denV gene of bacteriophage T4
- 1 April 1986
- journal article
- research article
- Published by Springer Nature in Molecular Genetics and Genomics
- Vol. 203 (1) , 163-171
- https://doi.org/10.1007/bf00330398
Abstract
The denV gene of bacteriophage T4 was reconstituted from two overlapping DNA fragments cloned in M13 vectors. The coding region of the intact gene was tailored into a series of plasmid vectors containing different promoters suitable for expression of the gene in E. coli and in yeast. Induction of the TAC promoter with IPTG resulted in overexpression of the gene, which was lethal to E. coli. Expression of the TACdenV gene in the absence of IPTG, or the use of the yeast GAL1 or ADH promoters resulted in partial complementation of the UV sensitivity of uvrA, uvrB, uvrC and recA mutants of E. coli and rad1, rad2, rad3, rad4 and rad10 mutants of S. cerevisiae. The extent of denV-mediated reactivation of excision-defective mutants was approximately equal to that of photoreactivation of such strains. Excision proficient E. coli cells transformed with a plasmid containing the denV gene were slightly more resistant to ultraviolet (UV) radiation than control cells without the denV gene. On the other hand, excision proficient yeast cells were slightly more sensitive to killing by UV radiation following transformation with a plasmid containing the denV gene. This effect was more pronounced in yeast mutants of the RAD52 epistasis group.Keywords
This publication has 46 references indexed in Scilit:
- Identification, physical map location and sequence of theden Vgene from bacteriophage T4Nucleic Acids Research, 1984
- DNA N-glycosylases and UV repairNature, 1980
- A pyrimidine dimer–DNA glycosylase activity associated with the v gene product of bacteriophage T4Nature, 1980
- Repair of DNA damaged by methyl methanesulfonate in bacteriophage T4Biochimica et Biophysica Acta (BBA) - Nucleic Acids and Protein Synthesis, 1976
- Further purification and characterization of T4 endonuclease VBiochimica et Biophysica Acta (BBA) - Nucleic Acids and Protein Synthesis, 1976
- REPAIR OF DNA DAMAGED BY 4-NITROQUINOLINE 1-OXIDE: A COMPARISON OF ESCHERICHIA COLI AND BACTERIOPHAGE T4 REPAIR SYSTEMSThe Japanese Journal of Genetics, 1976
- Nonsense mutants in the bacteriophage T4D v geneMutation Research - Fundamental and Molecular Mechanisms of Mutagenesis, 1975
- Studies on the substrate specificity of the T4 excision repair endonucleaseMutation Research - Fundamental and Molecular Mechanisms of Mutagenesis, 1972
- Cleavage of Structural Proteins during the Assembly of the Head of Bacteriophage T4Nature, 1970
- Recovery of UV-inactivated E. coli cells by the v-gene action of phage T4Mutation Research - Fundamental and Molecular Mechanisms of Mutagenesis, 1968