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The wild-type Bacillus subtilis spore protein, SspCwt, binds to DNA in vitro and in vivo and changes the conformation of DNA from B to A. Synthesis of the cloned SspCwt gene in Escherichia coli also causes large increases in mutation frequency. Binding of SspCwt to transforming DNA from Haemophilus influenzae made the DNA resistant to ultraviolet (UV) radiation. The mutant protein, SspCala, which does not bind DNA, did not change the UV resistance. The UV sensitivity of the DNA/SspCwt complex was not increased when the recipients of the DNA were defective in excision of pyrimidine dimers. These data indicate that the H. influenzae excision mechanism does not operate on the spore photoproduct formed by UV irradiation of the complex. Selection for the streptomycin- or erythromycin-resistance markers on the transforming DNA evidenced significant mutations at loci closely linked to these, but not at other loci. SspCwt apparently entered the cell attached to the transforming DNA, and caused mutations in adjacent loci. The amount of such mutations decreased when the transforming DNA was UV irradiated, because UV unlinks linked markers.
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p.107-114,fig.
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autor
- Brookhaven National Laboratory, Upton, NY 11973, USA
autor
autor
Bibliografia
- 1. Mohr, S.C., Sokolov, N.V.H.A.. He, C. & Setlow, P. (1991) Binding of small, acid-soluble spore proteins from Bacillus subtilis changes the conformation of DNA from B to A. Proc. Natl. Acad. Sri. U.S.A. 88,77-81.
- 2. Setlow, P. (ll>92) DNA in dormant spores of Bacillus species is in an A-like conformation. Mol. Microbiol, 6, 563-567.
- 3. Nicholson, W.L., Setlow, B. & Setlow, P. (1991) Ultraviolet irradiation of DNAcomplexed with a/p-type small, acid-soluble proteins from spores of Bacillus or Clostridium species makes spore photoproduct but not thymine dimers. Proc. Natl. Acad. Sci. U.S.A. 88,8288-8292.
- 4. Fairhead, H. & Setlow, P. (1992) Binding of DNA to a/p-type small, acid-soluble proteins from spores of Bacillus or Clostridium species prevents formation of cvtosine dimers. cytosine thymine dimers and bipyrimidine photoadducts upon ultraviolet irradiation. J. Bacterial. 174, 2874- -2880.
- 5. Setlow. B.. Hand, A.R. & Setlow, P. (1991) Synthesis of a Bacillus subtilis small, acid-soluble spore protein in Escherichia coli causes cell DNA to assume some characteristics of spore DNA. ]. Bacteriol. 173,1642-1653.
- 6. Setlow,J.K., Randesi, M., Adams,J.G., Setlow, B. & Setlow. P. (1992) Mutation and killing of Escherichia coli expressing a cloned Bacillus subtilis gene whose product alters DNA conformation. /. Bacteriol. 174, 2943-2950.
- 7. Steinhart, W.L. & Herriott, R.M. (1968) Fate of recipient deoxyribonucleic acid transformation in Haemophilus influenzae. J. Bacteriol. 96, 1718-1724.
- 8. Beattie, K.L. & Setlow, J.K. (1970) Repair of ultraviolet-irradiated transforming deoxyribonucleic acid in Haemophilus influenzae. ). Bacteriol. 101,808-812.
- 9. Setlow, J.K., Boling, M.E., Beattie, K.L. & Kimball. R.F. (1972) Acomplex of recombination and repair genes in Haemophilus influenzae. J. Mol. Biol. 68, 361-378.
- 10. LeClerc, J.E. & Setlow. J.K. (1973) Effects of combining ultraviolet repair and recombination mutations in Haemophilus influenzae. Nature Nru? Biol. 241, 172-174.
- 11. Marmur, J. (1961) A procedure for the isolation of deoxvribonucleic acid from microorganisms. J. Mol. Biol. 3,208-218.
- 12. Tovar-Rojo, F. & Setlow, P. (1991) Effects of mutant small, acid-soluble spore proteins from Bacillus subtilis on DNA in vivo and in vitro. /. Bacteriol. 173, 4827-4835.
- 13. Connors. M.J. & Setlow, P. (1985) Cloning of a small, acid-soluble spore protein from Bacillus subtilis and determination of its complete nucleotide sequence. /. Bacteriol. 161.333-339.
- 14. Setlow, B.. Sun. D. & Setlow. P. (1992) Studies of the interaction between DNA and a/p-type small, acid-soluble spore proteins: a new class of DNA binding protein. }. Bacteriol. 174, 2312-2322.
- 15. Kimball, R.F. & Setlow, J.K. (1972) Mutations induced in Haemophilus influenzae by transformation with nitrosoguanidine-treated DNA. Mutation Res. 14.137-146.
- 16. Setlow, B. & Setlow, P. (1993) Dipicolinic acid greatly enhances the production of spore photoproduct in bacterial spores upon ultraviolet irradiation. Appl. Env. Microbiol. 59. 640-643.
- 17. Riklis, E. (1965) Studies on mechanism of repair of ultraviolet-irradiated viral and bacterial DNA in vivo and in vitro. Can. /. Biochem. 43, 1207-1219.
- 18. Rahn, R.O. & Hosszu, J.L. (1969) Influence of relative humidity on the photochemistry of DNA films. Biochim. Biophys. Acta 190,126-131.
- 19. Setlow, J.K. (1977) The shape of the ultraviolet inactivation curve for transforming DNA. Nature (London) 268.169-170.
- 20. Herriott, R.M. (1971) Effects on DNA: transforming principle; in Chemical Mutagens (Hollaender, A.,ed.) vol. l.pp-173-217, Plenum Publishing Corp., New York.
- 21. Goodgal, S.H. (1961) Studies on transformation of Hemophilus influenzae. IV. Linked and unlinked transformations. J. Gen. Physiol. 45, 205-228.
- 22. Setlow, B. & Setlow, P. (1995) Binding to DNA protects a/3-type small, acid-soluble spore proteins of Bacillus and Clostridium species against digestion by their specific protease as well as by other proteases. J. Bacteriol. 177, 4149-4151.
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Bibliografia
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