Abstract
An enzyme activity, named T4 endonuclease V, was purified from T4-infected Escherichia coli. The enzyme induces single-stranded breaks in ultraviolet-irradiated DNA but does not act on native or heat-denatured DNA. The enzyme activity is dependent on the dose of ultraviolet irradiation, and the number of the breaks formed is approximately equal to the number of pyrimidine dimers present in the DNA. Denatured DNA, which has been exposed to ultraviolet light, is also attacked by the enzyme although the extent of the reaction is greater with irradiated native DNA. The enzyme shows optimal activity at pH 7.5 and does not require added divalent ions. When the enzyme-treated, irradiated DNA is subjected to stepwise degradation by spleen phosphodiesterase, dimers are released more rapidly than thymine into the acid-soluble fraction, suggesting that the enzyme induces a break at the 5′-side of a pyrimidine dimer. The enzyme, whose formation is controlled by the v+ gene of T4, appears to be responsible for the first step of excision repair.
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Selected References
These references are in PubMed. This may not be the complete list of references from this article.
- BOYCE R. P., HOWARD-FLANDERS P. RELEASE OF ULTRAVIOLET LIGHT-INDUCED THYMINE DIMERS FROM DNA IN E. COLI K-12. Proc Natl Acad Sci U S A. 1964 Feb;51:293–300. doi: 10.1073/pnas.51.2.293. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Carrier W. L., Setlow R. B. Endonuclease from Micrococcus luteus which has activity toward ultraviolet-irradiated deoxyribonucleic acid: purification and properties. J Bacteriol. 1970 Apr;102(1):178–186. doi: 10.1128/jb.102.1.178-186.1970. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Cleaver J. E. Defective repair replication of DNA in xeroderma pigmentosum. Nature. 1968 May 18;218(5142):652–656. doi: 10.1038/218652a0. [DOI] [PubMed] [Google Scholar]
- Dürwald H., Hoffmann-Berling H. Endonuclease-I-deficient and ribonuclease I-deficient Escherichia coli mutants. J Mol Biol. 1968 Jul 14;34(2):331–346. doi: 10.1016/0022-2836(68)90257-x. [DOI] [PubMed] [Google Scholar]
- Friedberg E. C., Goldthwait D. A. Endonuclease II of E. coli. I. Isolation and purification. Proc Natl Acad Sci U S A. 1969 Mar;62(3):934–940. doi: 10.1073/pnas.62.3.934. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Friedberg E. C., King J. J. Endonucleolytic cleavage of UV-irradiated DNA controlled by the V+ gene in phage T4. Biochem Biophys Res Commun. 1969 Nov 6;37(4):646–651. doi: 10.1016/0006-291x(69)90859-6. [DOI] [PubMed] [Google Scholar]
- Kaplan J. C., Kushner S. R., Grossman L. Enzymatic repair of DNA, 1. Purification of two enzymes involved in the excision of thymine dimers from ultraviolet-irradiated DNA. Proc Natl Acad Sci U S A. 1969 May;63(1):144–151. doi: 10.1073/pnas.63.1.144. [DOI] [PMC free article] [PubMed] [Google Scholar]
- LEHMAN I. R., ROUSSOS G. G., PRATT E. A. The deoxyribonucleases of Escherichia coli. II. Purification and properties of a ribonucleic acid-inhibitable endonuclease. J Biol Chem. 1962 Mar;237:819–828. [PubMed] [Google Scholar]
- Meselson M., Yuan R. DNA restriction enzyme from E. coli. Nature. 1968 Mar 23;217(5134):1110–1114. doi: 10.1038/2171110a0. [DOI] [PubMed] [Google Scholar]
- RAZZELL W. E., KHORANA H. G. Studies on polynucleotides. IV. Enzymic degradation; the stepwise action of venom phosphodiesterase on deoxyribo-oligonucleotides. J Biol Chem. 1959 Aug;234(8):2114–2117. [PubMed] [Google Scholar]
- RAZZELL W. E., KHORANA H. G. Studies on polynucleotides. X. Enzymic degradation. Some properties and mode of action of spleen phosphodiesterase. J Biol Chem. 1961 Apr;236:1144–1149. [PubMed] [Google Scholar]
- SETLOW R. B., CARRIER W. L. THE DISAPPEARANCE OF THYMINE DIMERS FROM DNA: AN ERROR-CORRECTING MECHANISM. Proc Natl Acad Sci U S A. 1964 Feb;51:226–231. doi: 10.1073/pnas.51.2.226. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Sadowski P. D., Hurwitz J. Enzymatic breakage of deoxyribonucleic acid. I. Purification and properties of endonuclease II from T4 phage-infected Escherichia coli. J Biol Chem. 1969 Nov 25;244(22):6182–6191. [PubMed] [Google Scholar]
- Sekiguchi M., Yasuda S., Okubo S., Nakayama H., Shimada K., Takagi Y. Mechanism of repair of DNA in bacteriophage. I. Excision of pyrimidine dimers from ultraviolet-irradiated DNA by an extract of T4-infected cells. J Mol Biol. 1970 Jan 28;47(2):231–242. doi: 10.1016/0022-2836(70)90342-6. [DOI] [PubMed] [Google Scholar]
- Takagi Y., Sekiguchi M., Okubo S., Nakayama H., Shimada K., Yasuda S., Nishimoto T., Yoshihara H. Nucleases specific for ultraviolet light-irradiated DNA and their possible role in dark repair. Cold Spring Harb Symp Quant Biol. 1968;33:219–227. doi: 10.1101/sqb.1968.033.01.025. [DOI] [PubMed] [Google Scholar]
- Weiss B., Live T. R., Richardson C. C. Enzymatic breakage and joining of deoxyribonucleic acid. V. End group labeling and analysis of deoxyribonucleic acid containing single straned breaks. J Biol Chem. 1968 Sep 10;243(17):4530–4542. [PubMed] [Google Scholar]
- Yasuda S., Sekiguchi M. Mechanism of repair of DNA in bacteriophage. II. Inability of ultraviolet-sensitive strains of bacteriophage in inducing an enzyme activity to excise pyrimidine dimers. J Mol Biol. 1970 Jan 28;47(2):243–255. doi: 10.1016/0022-2836(70)90343-8. [DOI] [PubMed] [Google Scholar]