Summary
The sequences of a collection of 261 spontaneous lacI- mutants recovered in a PolA- strain of Escherichia coli have indicated an increase in the frequency of most classes of mutation in this strain. Among base substitutions in lacI, a preference for transversions over transitions was observed. In addition, a single transition in the lac operator was enhanced 8-fold. More significantly, of 18 frameshifts, 12 occurred adjacent to a 5′-GTGG-3′ sequence. Likewise, 15 of 24 deletions and 2 of 10 duplications had 5′-GTGG-3′ sequences at one or both endpoints. We speculate that the prevalence of mutations at these specific sequences reflects the persistence of strand discontinuities that enhance the opportunity for mutagenic mishaps. Further, 5′-GTGG-3′ sequences apparently represent sites where DNA polymerase I is involved in some aspect of DNA metabolism. These results strengthen the view that DNA context contributes an important component to spontaneous mutagenesis and indicate an anti-mutagenic role for DNA polymerase I.
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References
Albertini AM, Hofer 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
Cheung S, Arndt K, Lu P (1984) Correlation of lac operator DNA imino proton exchange kinetics with its function. Proc Natl Acad Sci USA 81:3665–3669
Coukell MB, Yanovsky C (1970) Increased frequency of deletions in DNA polymerase mutants of Escherichia coli. Nature 228:633–636
Coulondre C, Miller JH, Farabaugh PJ, Gilbert W (1978) Molecular basis of base substitution hotspots in Escherichia coli. Nature 274:775–780
De Lucia P, Cairns J (1969) Isolation of an E. coli strain with a mutation affecting DNA polymerase. Nature 224:1164–1166
Drake JW (1970) The molecular basis of mutation. Holden Day, San Francisco, pp 177–185
Drake JW, Allen EF, Forsberg SA, Preparata RM, Greening EO (1969) Spontaneous mutation: genetic control of mutations rates in bacteriophage T4. Nature 221:1128–1132
Farabaugh PJ (1978) Sequence of the lacI gene. Nature 274:765–769
Farabaugh PJ, Schmeissner U, Hofer M, Miller JH (1978) Genetic studies of the lac repressor. VII. On the molecular basis of spontaneous hotspots in the lacI gene of Escherichia coli. J Mol Biol 126:847–863
Fowler RG, Schaaper RM, Glickman BW (1986) Characterization of mutational specificity within the lacI gene for a mutD5 mutator strain of Escherichia coli defective in 3′→5′ exonuclease (proofreading) activity. J Bacteriol 167:130–137
Fuller RS, Funnell BE, Kornberg A (1984) The dnaA protein complex with the E. coli chromosomal replication origin (oriC) and other DNA sites. Cell 38:889–900
Glickman BW, Ripley LS (1984) Structural intermediates of deletion mutagenesis: a role for palindromic DNA. Proc Natl Acad Sci USA 81:512–516
Glickman BW, van der Elsen P, Radman M (1978) Induced mutagenesis in dam - mutants of Escherichia coli: a role for 6-methyl adenine residues in mutation avoidance. Mol Gen Genet 163:307–312
Glickman BW, Burns PA, Fix DF (1986) Mechanisms of spontaneous mutagenesis: clues from altered mutational specificity in DNA repair defective strains. In: Shankel DM, Hartman PE, Kada T, Hollaender A (eds) Antimutagenesis and Anticarcinogenesis, Mechanisms, Plenum Press, New York, pp 259–281
Gross J, Gross M (1969) Genetic analysis of an E. coli strain with a mutation affecting DNA polymerase Nature 224:1166–1168
Hermbomel P, Bourachot B, Yaniv M (1984) Two distinct enhancers with different cell specificities coexist in the regulatory region of polyoma. Cell 39:653–662
Joyce CM, Fujii DM, Laks HS, Hughes CM, Grindley NDF (1985) Genetic mapping and DNA sequence analysis of mutations in the polA gene of Escherichia coli. J Mol Biol 186:283–293
Konrad EB (1978) Isolation of an Escherichia coli K-12 dnaE mutation as a mutator. J Bacteriol 133:1197–1202
Kornberg A (1980) DNA Replication. Freeman, San Francisco, CA
Kunkel TA (1984) The mutational specificity of depurination. Proc Natl Acad Sci USA 81:1494–1498
Kunz BA, Glickman BW (1985) Mechanism of mutation by thymine starvation in Escherichia coli: clues from mutagenic specificity. J Bacteriol 162:859–864
Lockshon D, Morris DR (1985) Sites of reaction of Escherichia coli DNA gyrase on pBR322 in vivo as revealed by oxolinic acid-induced plasmid linearization. J Mol Biol 181:63–74
Lu AL, Welsh K, Clark S, Su SS, Modrich P (1984) Repair of DNA basepair mismatches in extracts of Escherichia coli. Cold Spring Harbor Symp Quant Biol 49:589–596
Ripley LS (1982) Model for the participation of quasipalindromic DNA sequences in frameshift mutation. Proc Natl Acad Sci USA 79:4128–4132
Ripley LS, Glickman BW (1982) The unique self-complementarity of palindromic sequences provides DNA structural intermediates for mutation. Cold Spring Harbor Symp Quant Biol 47:851–861
Sanger F, Nicklen S, Coulson AR (1977) DNA sequencing with chainterminating inhibitors. Proc Natl Acad Sci USA 74:5463–5467
Sargentini NJ, Smith KC (1981) Much of spontaneous mutagenesis in Escherichia coli is due to error-prone DNA repair: implications for spontaneous carcinogenesis. Carcinogenesis 2:863–872
Sargentini NJ, Smith KC (1985) Spontaneous mutagenesis: the roles of DNA repair, replication and recombination. Mutat Res 154:1–27
Savic DJ, Romac SP (1982) Powerful mutator activity of the polA1 mutation within the histidine region of Escherichia coli K-12. J Bacteriol 149:955–960
Schaaper RM, Danforth BN, Glickman BW (1985) Rapid repeated cloning of mutant lac repressor genes. Gene 39:181–189
Schaaper RM, Danforth BN, Glickman BW (1986) Mechanisms of spontaneous mutagenesis: an analysis of the spectrum of spontaneous mutation in the Escherichia coli lacI gene. J Mol Biol 189:273–284
Schmiessner U, Ganem D, Miller JH (1977) Genetic studies of the lac repressor. II. Fine structure deletion map of the lacI gene and its correlation with the physical map. J Mol Biol 109:303–326
Siegel EC, Vaccaro KK (1978) The reversion of trp frameshift mutations in mut, polA, lig and dnaE mutant strains of Escherichia coli. Mutat Res 50:9–17
Smith GR (1983) Chi hotspots of generalized recombination. Cell 34:709–710
Speyer JF, Karam JD, Lenny AB (1966) On the role of DNA polymerase in base selection. Cold Spring Harbor Symp Quant Biol 31:693–697
Streisinger G, Okada Y, Emrich J, Newton J, Tsugita A, Terzaghi E, Inouye M (1966) Frameshift mutations and the genetic code. Cold Spring Harbor Symp Quant Biol 33:77–84
Topal MD, Fresco JR (1976) Complementary base pairing and the origin of substitution mutations. Nature 263:285–289
Vaccaro KK, Siegel EC (1975) Increased spontaneous reversion of certain frameshift mutations in DNA polymerase I deficient strains of Escherichia coli. Mol Gen Genet 141:251–289
Vogel HJ, Bonner DM (1956) Acetylorithinase of Escherichia coli: partial purification and some properties. J Biol Chem 218:97–106
von Borstel RC (1969) On the origin of spontaneous mutations. Jpn J Genet 44:102–105
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Fix, D.F., Burns, P.A. & Glickman, B.W. DNA sequence analysis of spontaneous mutation in a PolA1 strain of Escherichia coli indicates sequence-specific effects. Mol Gen Genet 207, 267–272 (1987). https://doi.org/10.1007/BF00331588
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DOI: https://doi.org/10.1007/BF00331588