Summary
Chromosomal rearrangements, uniformly represented by very large deletions, were stimulated upon transiently exposing Escherichia coli cells with a defective lambda prophage to about 18% (v/v) ethanol. It was shown that the ethanol treatment induced deletion formation rather than enriching for ethanol-tolerant cells. The deletions in 435 mutants were mapped to 26 groups. Ethanol treatment changed the spectrum of deletions relative to those arising spontaneously, and stimulated the formation of deletions with endpoints in E. coli DNA flanking the lambda fragment. The promotion of deletion formation by ethanol involves the joining of distant, nonhomologous linear DNA segments, which can be considered an illegitimate recombination event; however, activity of the E. coli recA gene product was also required. Although spontaneous deletions arose in comparable cells defective for recA, the incidence of deletion formation in recA cells was not altered by ethanol. It is proposed that ethanol stimulates chromosomal rearrangements involving two oppositely oriented replication forks, since the localized deletions commonly removed or inactivated the chromosomal segment including the bidirectional lambda origin of replication. The results imply a novel mutagenic process induced by an agent that does not act directly on DNA.
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References
Adhya S, Cleary P, Campbell A (1968) A deletion analysis of prophage lambda and adjacent regions. Proc Natl Acad Sci USA 61:956–962
Albertini AM, Hoefer M, Calos MP, Miller JH (1982) On the formation of spontaneous deletions: The importance of short sequence homologies in the generation of large deletions. Cell 29:319–328
Allgood ND, Silhavy TJ (1988) Illegitimate recombination in bacteria. In: Kucherlapati R, Smith GR (eds) Genetic recombination. ASM Publications, Washington, pp 309–330
Arber W, Enquist L, Hohn B, Murray NE, Murray K (1983) Experimental methods for use with lambda. In: Hendrix RW, Roberts JW, Stahl FW, Weisberg RA (eds) Lambda II. Cold Spring Harbor Laboratory, Cold Spring Harbor, NY, pp 433–466
Bachmann BJ (1987) Derivations and genotypes of some mutant derivatives of Escherichia coli K-12. In: Neidhardt FC, Ingraham JL, Low KB, Magasanik B, Schaechter M, Umbarger HE (eds) Escherichia coli and Salmonella typhimurium: Cellular and molecular biology. ASM Publications, Washington, pp 1190–1219
Bachmann BJ (1990) Linkage map of Escherichia coli, edn 8. Microbiol Rev 54:130–197
Bochner BR, Lee PC, Wilson SW, Cutler CW, Ames BN (1984) AppppA and related adenylated nucleotides are synthesized as a consequence of oxidation stress. Cell 37:225–232
Cashell M, Rudd KE (1987) The stringent response. In: Neidhardt FC, Ingraham JL, Low KB, Magasanik B, Schaechter M, Umbarger HE (eds) Escherichia coli and Salmonella typhimurium: Cellular and molecular biology. ASM Publications, Washington, pp 1410–1438
Daniels DL, Schroder JL, Szybalski W, Sanger F, Coulson AR, Hong GF, Hill DF, Peterson GB, Blattner FR (1983) Complete annotated lambda sequence. In: Hendrix RW, Roberts JW, Stahl FW, Weisberg RA (eds) Lambda II. Cold Spring Harbor Laboratory, Cold Spring Harbor, NY, pp 519–676
Davis RW, Parkinson JS (1971) Deletion mutants of bacteriophage lambda. III. Physical structure of att. J Mol Biol 56:403–423
Enquist L, Skalka A (1973) Replication of bacteriophage λ DNA dependent on the function of host and viral genes. I. Interaction of red, gam, and rec. J Mol Biol 75:185–212
Flamm EL, Weisberg RA (1985) Primary structure of the hip gene of Escherichia coli and of its product, the beta-subunit of integration host factor. J Mol Biol 170:611–633
Franklin NC (1967) Extraordinary recombinational events in Escherichia coli. Their independence of the rec + function. Genetics 55:699–707
Franklin NC (1971) Illegitimate recombination. In: Hershey AD (ed) The bacteriophage lambda. Cold Spring Harbor Laboratory, Cold Spring Harbor, NY, pp 175–194
Hayes S (1985) Ethanol-induced genotoxicity. Mutat Res 143:23–27
Hayes S (1988) Mutations suppressing the loss of replication control: genetic analysis of λ dependent replicative-killing, replication initiation, and mutagenic mechanism. In: Moses RE, Summers WC (eds) DNA replication and mutagenesis, ASM Publications, Washington, pp 367–377
Hayes S, Hayes C (1986) Spontaneous λ o R mutations suppress inhibition of bacteriophage growth by nonimmune exclusion phenotype of defective λ prophage. J Virol 58:835–842
Hayes S, Duncan D, Hayes C (1990a) Alcohol treatment of defective lambda lysogens is deletionogenic. Mol Gen Genet 222:17–24
Hayes S, Hayes C, Duncan D, Bennett V, Blushke J (1990b) Stimulation of mutations suppressing the loss of replication control by small alcohols. Mutat Res 231:151–163
Höög J-O, Bahr-Lindström H von, Jörnvall H, Holmgren A (1986) Cloning and expression of the glutaredoxin (grx) gene of Escherichia coli. Gene 43:13–21
Landy A, Ross W (1977) Viral integration and excision: Structure of the lambda att sites. Science 197:1147–1159
Lee PC, Bochner BR, Ames BN (1983) AppppA, heat-shock stress, and cell oxidation. Proc Natl Acad Sci USA 80:7496–7500
Masters PS, Hong J-S (1981) Genetics of glutamine transport system in Escherichia coli. J Bacteriol 147:805–819
Miller JK, Barnes WM (1986) Colony probing as an alternative to standard sequencing as a means of direct analysis of chromosomal DNA to determine the spectrum of single base changes in regions of known sequence. Proc Natl Acad Sci USA 83:1026–1030
Mosig G (1988) Mapping and map distortions in bacteriophage crosses. In: Kucherlapati R, Smith GR (eds) Genetic recombination. ASM Publications, Washington, pp 141–167
Nohno T, Saito T, Hong J-S (1986) Cloning and complete nucleotide sequence of the Escherichia coli glutamate permease operon (glnHPQ). Mol Gen Genet 205:260–269
Radding CM (1988) Homologous pairing and strand exchange promoted by Escherichia coli RecA protein. In: Kucherlapati R, Smith GR (eds) Genetic recombination. ASM Publications, Washington, pp 193–229
Roberts JW, Devoret R (1983) Lysogenic induction. In: Hendrix RW, Roberts JW, Stahl FW, Weisberg RA (eds) Lambda II. Cold Spring Harbor Laboratory, Cold Spring Harbor, NY, pp 123–144
Russel M, Holmgren A (1988) Construction and characterization of glutaredoxin-negative mutants of Escherichia coli. Proc Natl Acad Sci USA 85:990–994
Sato K, Campbell A (1970) Specialized transduction of galactose by lambda phage from a deletion lysogen. Virology 41:474–487
Shimada K, Weisberg RA, Gottesman ME (1972) Prophage lambda at unusual chromosomal locations. I. Location of the secondary attachment sites and the properties of the lysogens. J Mol Biol 63:483–503
Smith GR (1983) General recombination. In: Hendrix RW, Roberts JW, Stahl FW, Weisberg RA (eds) Lambda II. Cold Spring Harbor Laboratory, Cold Spring Harbor, NY, pp 175–209
Taylor AW (1988) RecBCD enzyme of Escherichia coli. In: Kucherlapati R, Smith GR (eds) Genetic recombination. ASM Publications, Washington, pp 231–263
Van Bogelen RA, Kelley PM, Neidhardt FC (1987) Differential induction of heat shock, SOS, and oxidation stress regulons and accumulation of nucleotides in Escherichia coli. J Bacteriol 169:26–32
Walker GC (1984) Mutagenesis and inducible responses to deoxyribonucleic acid damage in Escherichia coli. Microbiol Rev 48:60–93
Weisberg RA, Adhya S (1977) Illegitimate recombination in bacteria and bacteriophage. Annu Rev Genet 11:451–473
Zissler J, Signer ER, Schaefer F (1971) The role of recombination in growth of bacteriophage lambda. The gamma gene. In: Hershey A (ed) The bacteriophage lambda. Cold Spring Harbor Laboratory, Cold Spring Harbor, NY, pp 455–475
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Hayes, S. Mapping ethanol-induced deletions. Molec. Gen. Genet. 231, 139–149 (1991). https://doi.org/10.1007/BF00293831
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DOI: https://doi.org/10.1007/BF00293831