Enzymes involved in thymine dimer excision in bacteriophage T4-infected Escherichia coli

Pawl, G.; Taylor, Russell E.; Minton, Kenneth W.; Friedberg, Errol C.

doi:10.1016/s0022-2836(76)80097-6

Cited by 19 publications

(10 citation statements)

References 28 publications

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“…This product is encoded by the denV (DNA endonuclease V) gene (41) of phage T4. Mutants defective in this gene have the phenotype of abnormal UV sensitivity (15) and are defective in the excision of thymine-containing PD from their own DNA after infection of E. coli with UV-irradiated phage (27).…”

Section: Ie00 Kmentioning

confidence: 99%

den V gene of bacteriophage T4 codes for both pyrimidine dimer-DNA glycosylase and apyrimidinic endonuclease activities

McMillan

Edenberg

Radany

et al. 1981

J Virol

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Recent studies have shown purified preparations of phage T4 UV DNA-incising activity (T4 UV endonuclease or endonuclease V of phage T4) contain a pyrimidine dimer-DNA glycosylase activity that catalyzes hydrolysis of the 5' glycosyl bond of dimerized pyrimidines in UV-irradiated DNA. Such enzyme preparations have also been shown to catalyze the hydrolysis of phosphodiester bonds in UV-irradiated DNA at a neutral pH, presumably reflecting the action of an apurinic/apyrimidinic endonuclease at the apyrimidinic sites created by the pyrimidine dimer-DNA glycosylase. In this study we found that preparations of T4 UV DNA-incising activity contained apurinic/apyrimidinic endonuclease activity that nicked depurinated form I simian virus 40 DNA. Apurinic/apyrimidinic endonuclease activity was also found in extracts of Escherichia coli infected with T4 denV+ phage. Extracts of cells infected with T4 denV mutants contained significantly lower levels of apurinic/apyrimidinic endonuclease activity; these levels were no greater than the levels present in extracts of uninfected cells. Furthermore, the addition of DNA containing apurinic or apyrimidinic sites to reactions containing UV-irradiated DNA and T4 enzyme resulted in competition for pyrimidine dimer-DNA glycosylase activity against the UV-irradiated DNA. On the basis of these results, we concluded that apurinic/apyrimidinic endonuclease activity is encoded by the denV gene of phage T4, the same gene that codes for pyrimidine dimer-DNA glycosylase activity.

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Section: Ie00 Kmentioning

confidence: 99%

den V gene of bacteriophage T4 codes for both pyrimidine dimer-DNA glycosylase and apyrimidinic endonuclease activities

McMillan

Edenberg

Radany

et al. 1981

J Virol

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“…Photobiol., p. 318, 1980). After incision, the 5'-to-3' exonuclease activity of Escherichia coli polymerase I excises a majority of the dimers (17). E. coli polymerase I and phage ligase are required for resynthesis and sealing in the repaired region to form viable phage DNA (15).…”

mentioning

confidence: 99%

Excision repair and patch size in UV-irradiated bacteriophage T4

Yarosh¹,

Rosenstein²,

Setlow³

1981

J Virol

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We determined the average size of excision repair patches in repair of UV lesions in bacteriophage T4 by measuring the photolysis of bromodeoxyuridine incorporated during repair. The average patch was small, approximately four nucleotides long. In control experiments with the denV1 excision-deficient mutant, we encountered an artifact, a protein(s) which remained bound to phenol-extracted DNA and prevented nicking by the UV-specific endonucleases of Micrococcus luteus and bacteriophage T4.

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“…If the dimers are removed as oligomers, the dimer content of the high-molecular-weight component recovered from the gel would be markedly lower than that of the unfractionated sample. If, instead, the unexcised dimers remained associated with high-molecular-weight DNA, the thymine dimer content of the high-molecular-weight DNA recovered from the column should be approximately equivalent to that of the corresponding unfractionated sample (23). To differentiate between these two possibilities, exponentially growing 3H-labeled uvrD bacteria were irradiated with a dose of UV sufficient to yield on August 5, 2020 by guest http://jb.asm.org/ Downloaded from approximately 0.1% thymine as dimers and incubated under growth conditions for 0, 15, or 30 min.…”

Section: Resultsmentioning

confidence: 99%

“…Gel filtration of phenol-extracted samples was carried out on a Sephadex G75 (Pharmacia) column (0.9 by 22 cm) equilibrated with 10 mM Tris.hydrochloride (pH 8.0)-10 mM EDTA at 4°C as described by Pawl et al (23). Fractions of 1.2 ml were eluted with 10 mM Tris-hydrochloride (pH 8.0)-10 mM EDTA at a flow rate of 0.3 ml/min.…”

Section: Materlals and Methodsmentioning

confidence: 99%

Effect of the uvrD mutation on excision repair

Kuemmerle¹,

Masker²

1980

J Bacteriol

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A pair of related Escherichia coli K-12 strains, one of which contains the uvrD101 mutation, were constructed and compared for ability to perform various steps in the excision repair of deoxyribonucleic acid damage inflicted by ultraviolet radiation. The results of this study indicated: (i) ultraviolet sensitivity in the uvrD101 mutant was greater than that of wild type but less than that measured in an incision-deficient uvrA mutant; (ii) host cell reactivation paralleled the survival data; (iii) postirradiation deoxyribonucleic acid degradation was virtually identical in the two strains; (iv) incision, presumably at the sites of pyrimidine dimers, proceeded normally in the uvrD101 strain; (v) excision of pyrimidine dimers was markedly reduced in both rate and extent in the uvrD101 mutant; (vi) the amount of repair resynthesis was the same in both strains, and there was no evidence of abnormally long repair patches in the uvrD mutant; and (vii) rejoining of incision breaks was slow and incomplete in the uvrD strain. These data suggest that the ultraviolet sensitivity conferred by the uvrD mutation arises from inefficient removal of pyrimidine dimers or from failure to close incision breaks. The data are compatible with the notion that the uvrD+ gene produce affects the conformation of incised deoxyribonucleic acid molecules.

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Enzymes involved in thymine dimer excision in bacteriophage T4-infected Escherichia coli

Cited by 19 publications

References 28 publications

den V gene of bacteriophage T4 codes for both pyrimidine dimer-DNA glycosylase and apyrimidinic endonuclease activities

den V gene of bacteriophage T4 codes for both pyrimidine dimer-DNA glycosylase and apyrimidinic endonuclease activities

Excision repair and patch size in UV-irradiated bacteriophage T4

Effect of the uvrD mutation on excision repair

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