Genetics, Vol 125, 275-280, Copyright © 1990

INVESTIGATIONS

A Set of lacZ Mutations in Escherichia coli That Allow Rapid Detection of Specific Frameshift Mutations

C. G. Cupples, M. Cabrera, C. Cruz and J. H. Miller
Current address: Department of Biology, Concordia University, 1455 de Maisonmeuve Boulevard West, Montreal, PQ, H3G 1M8, Canada.

We have used site-directed mutagenesis to alter bases in lacZ near the region encoding essential residues in the active site of {beta}-galactosidase. The altered sequences generate runs of six or seven identical base pairs which create a frameshift, resulting in a Lac(-) phenotype. Reversion to Lac(+) in each strain can occur only by a specific frameshift at these sequences. Monotonous runs of A's (or of T's on the opposite strand) and G's (or C's) have been constructed, as has an alternating -C-G-sequence. These specific frameshift indicator strains complement a set of six previously described strains which detect each of the base substitutions. We have examined a variety of mutagens and mutators for their ability to cause reversion to Lac(+). Surprisingly, frameshifts are well stimulated at many of these runs by ethyl methanesulfonate, N-methyl-N'-nitro-N-nitrosoguanidine and 2-aminopurine, mutagens not widely known to induce frameshifts. A comparison of ethyl methanesulfonate, N-methyl-N'-nitro-N-nitrosoguanidine and 2-aminopurine frameshift specificity with that found with a mutH strain suggests that these mutagens partially or fully saturate or inactivate the methylation-directed mismatch repair system and allow replication errors leading to frameshifts to escape repair. This results in a form of indirect mutagenesis, which can be detected at certain sites.

This article has been cited by other articles:

				G. Van der Auwera, J. Baute, M. Bauwens, I. Peck, D. Piette, M. Pycke, P. Asselman, and A. Depicker Development and Application of Novel Constructs to Score C:G-to-T:A Transitions and Homologous Recombination in Arabidopsis Plant Physiology, January 1, 2008; 146(1): 22 - 31. [Abstract] [Full Text] [PDF]

				L. H. Sanders, A. Rockel, H. Lu, D. J. Wozniak, and M. D. Sutton Role of Pseudomonas aeruginosa dinB-Encoded DNA Polymerase IV in Mutagenesis J. Bacteriol., December 15, 2006; 188(24): 8573 - 8585. [Abstract] [Full Text] [PDF]

				B. Budke and A. Kuzminov Hypoxanthine Incorporation Is Nonmutagenic in Escherichia coli. J. Bacteriol., September 1, 2006; 188(18): 6553 - 6560. [Abstract] [Full Text] [PDF]

				L.L. Souza, I.R. Eduardo, M. Padula, and A.C. Leitao Endonuclease IV and Exonuclease III are involved in the repair and mutagenesis of DNA lesions induced by UVB in Escherichia coli Mutagenesis, March 1, 2006; 21(2): 125 - 130. [Abstract] [Full Text] [PDF]

				J.-P. Gratia Noncomplementing diploidy resulting from spontaneous zygogenesis in Escherichia coli Microbiology, September 1, 2005; 151(9): 2947 - 2959. [Abstract] [Full Text] [PDF]

				V. Gabrovsky, M. L. Yamamoto, and J. H. Miller Mutator Effects in Escherichia coli Caused by the Expression of Specific Foreign Genes J. Bacteriol., July 15, 2005; 187(14): 5044 - 5048. [Abstract] [Full Text] [PDF]

				T. Shibata, T. Hishida, Y. Kubota, Y.-W. Han, H. Iwasaki, and H. Shinagawa Functional overlap between RecA and MgsA (RarA) in the rescue of stalled replication forks in Escherichia coli Genes Cells, March 1, 2005; 10(3): 181 - 191. [Abstract] [Full Text] [PDF]

				E. S. Hong, A. Yeung, P. Funchain, M. M. Slupska, and J. H. Miller Mutants with Temperature-Sensitive Defects in the Escherichia coli Mismatch Repair System: Sensitivity to Mispairs Generated In Vivo J. Bacteriol., February 1, 2005; 187(3): 840 - 846. [Abstract] [Full Text] [PDF]

				W. Kuban, P. Jonczyk, D. Gawel, K. Malanowska, R. M. Schaaper, and I. J. Fijalkowska Role of Escherichia coli DNA Polymerase IV in In Vivo Replication Fidelity J. Bacteriol., July 15, 2004; 186(14): 4802 - 4807. [Abstract] [Full Text] [PDF]

				S. A. Taft-Benz and R. M. Schaaper The {theta} Subunit of Escherichia coli DNA Polymerase III: a Role in Stabilizing the {varepsilon} Proofreading Subunit J. Bacteriol., May 1, 2004; 186(9): 2774 - 2780. [Abstract] [Full Text] [PDF]

				E. Wolff, M. Kim, K. Hu, H. Yang, and J. H. Miller Polymerases Leave Fingerprints: Analysis of the Mutational Spectrum in Escherichia coli rpoB To Assess the Role of Polymerase IV in Spontaneous Mutation J. Bacteriol., May 1, 2004; 186(9): 2900 - 2905. [Abstract] [Full Text] [PDF]

				J.-P. Gratia and M. Thiry Spontaneous zygogenesis in Escherichia coli, a form of true sexuality in prokaryotes Microbiology, September 1, 2003; 149(9): 2571 - 2584. [Abstract] [Full Text] [PDF]

				J. M. Leonard, S. R. Bollmann, and J. B. Hays Reduction of Stability of Arabidopsis Genomic and Transgenic DNA-Repeat Sequences (Microsatellites) by Inactivation of AtMSH2 Mismatch-Repair Function Plant Physiology, September 1, 2003; 133(1): 328 - 338. [Abstract] [Full Text] [PDF]

				I. Bjedov, O. Tenaillon, B. Gerard, V. Souza, E. Denamur, M. Radman, F. Taddei, and I. Matic Stress-Induced Mutagenesis in Bacteria Science, May 30, 2003; 300(5624): 1404 - 1409. [Abstract] [Full Text] [PDF]

				I. Matic, A. Babic, and M. Radman 2-Aminopurine Allows Interspecies Recombination by a Reversible Inactivation of the Escherichia coli Mismatch Repair System J. Bacteriol., February 15, 2003; 185(4): 1459 - 1461. [Abstract] [Full Text] [PDF]

				A. A. M. Al Mamun, K. J. Marians, and M. Z. Humayun DNA Polymerase III from Escherichia coli Cells Expressing mutA Mistranslator tRNA Is Error-prone J. Biol. Chem., November 22, 2002; 277(48): 46319 - 46327. [Abstract] [Full Text] [PDF]

				G. R. Hoffmann, D. J. Crowley, and P. J. Theophiles Comparative potencies of induction of point mutations and genetic duplications by the methylating agents methylazoxymethanol and dimethyl sulfate in bacteria Mutagenesis, September 1, 2002; 17(5): 439 - 444. [Abstract] [Full Text] [PDF]

				J. H. Miller, P. Funchain, W. Clendenin, T. Huang, A. Nguyen, E. Wolff, A. Yeung, J.-H. Chiang, L. Garibyan, M. M. Slupska, et al. Escherichia coli Strains (ndk) Lacking Nucleoside Diphosphate Kinase Are Powerful Mutators for Base Substitutions and Frameshifts in Mismatch-Repair-Deficient Strains Genetics, September 1, 2002; 162(1): 5 - 13. [Abstract] [Full Text] [PDF]

				D. Gawel, M. Maliszewska-Tkaczyk, P. Jonczyk, R. M. Schaaper, and I. J. Fijalkowska Lack of Strand Bias in UV-Induced Mutagenesis in Escherichia coli J. Bacteriol., August 15, 2002; 184(16): 4449 - 4454. [Abstract] [Full Text] [PDF]

				K. Negishi, D. Loakes, and R. M. Schaaper Saturation of DNA Mismatch Repair and Error Catastrophe by a Base Analogue in Escherichia coli Genetics, August 1, 2002; 161(4): 1363 - 1371. [Abstract] [Full Text] [PDF]

				S. T. Fitz-Gibbon, H. Ladner, U.-J. Kim, K. O. Stetter, M. I. Simon, and J. H. Miller Genome sequence of the hyperthermophilic crenarchaeon Pyrobaculum aerophilum PNAS, January 9, 2002; (2002) 241636498. [Abstract] [Full Text] [PDF]

				D. C. Bell and C. G. Cupples Very-Short-Patch Repair in Escherichia coli Requires the dam Adenine Methylase J. Bacteriol., June 15, 2001; 183(12): 3631 - 3635. [Abstract] [Full Text]

				P. Funchain, A. Yeung, J. Stewart, W. M. Clendenin, and J. H. Miller Amplification of Mutator Cells in a Population as a Result of Horizontal Transfer J. Bacteriol., June 15, 2001; 183(12): 3737 - 3741. [Abstract] [Full Text]

				G. Macintyre, C. V. Atwood, and C. G. Cupples Lowering S-Adenosylmethionine Levels in Escherichia coli Modulates C-to-T Transition Mutations J. Bacteriol., February 1, 2001; 183(3): 921 - 927. [Abstract] [Full Text]

				B. S. Strauss, R. Roberts, L. Francis, and P. Pouryazdanparast Role of the dinB Gene Product in Spontaneous Mutation in Escherichia coli with an Impaired Replicative Polymerase J. Bacteriol., December 1, 2000; 182(23): 6742 - 6750. [Abstract] [Full Text]

				J. Zhao and M. E. Winkler Reduction of GC right-arrow TA Transversion Mutation by Overexpression of MutS in Escherichia coli K-12 J. Bacteriol., September 1, 2000; 182(17): 5025 - 5028. [Abstract] [Full Text]

				P. Funchain, A. Yeung, J. L. Stewart, R. Lin, M. M. Slupska, and J. H. Miller The Consequences of Growth of a Mutator Strain of Escherichia coli as Measured by Loss of Function Among Multiple Gene Targets and Loss of Fitness Genetics, March 1, 2000; 154(3): 959 - 970. [Abstract] [Full Text]

				H. Liu, S. R. Hewitt, and J. B. Hays Antagonism of Ultraviolet-Light Mutagenesis by the Methyl-Directed Mismatch-Repair System of Escherichia coli Genetics, February 1, 2000; 154(2): 503 - 512. [Abstract] [Full Text]

				J. O. Blaisdell, Z. Hatahet, and S. S. Wallace A Novel Role for Escherichia coli Endonuclease VIII in Prevention of Spontaneous Gright-arrowT Transversions J. Bacteriol., October 15, 1999; 181(20): 6396 - 6402. [Abstract] [Full Text]

				J. H. Miller, A. Suthar, J. Tai, A. Yeung, C. Truong, and J. L. Stewart Direct Selection for Mutators in Escherichia coli J. Bacteriol., March 1, 1999; 181(5): 1576 - 1584. [Abstract] [Full Text]

				M. C. Earley and G. F. Crouse The role of mismatch repair in the prevention of base pair mutations in Saccharomyces cerevisiae PNAS, December 22, 1998; 95(26): 15487 - 15491. [Abstract] [Full Text] [PDF]

				F. P. Guengerich, N. A. Hosea, A. Parikh, L. C. Bell-Parikh, W. W. Johnson, E. M. J. Gillam, and T. Shimada Twenty Years of Biochemistry of Human P450s. Purification, Expression, Mechanism, and Relevance to Drugs Drug Metab. Dispos., December 1, 1998; 26(12): 1175 - 1178. [Abstract] [Full Text]

				A. L. Jackson, R. Chen, and L. A. Loeb Induction of microsatellite instability by oxidative DNA damage PNAS, October 13, 1998; 95(21): 12468 - 12473. [Abstract] [Full Text] [PDF]

				M. Viswanathan and S. T. Lovett Single-Strand DNA-Specific Exonucleases in Escherichia coli: Roles in Repair and Mutation Avoidance Genetics, May 1, 1998; 149(1): 7 - 16. [Abstract] [Full Text] [PDF]

				S.-R. Kim, G. Maenhaut-Michel, M. Yamada, Y. Yamamoto, K. Matsui, T. Sofuni, T. Nohmi, and H. Ohmori Multiple pathways for SOS-induced mutagenesis in Escherichia coli: An overexpression of dinB/dinP results in strongly enhancing mutagenesis in the absence of any exogenous treatment to damage DNA PNAS, December 9, 1997; 94(25): 13792 - 13797. [Abstract] [Full Text] [PDF]

				R. S. Harris, G. Feng, K. J. Ross, R. Sidhu, C. Thulin, S. Longerich, S. K. Szigety, M. E. Winkler, and S. M. Rosenberg Mismatch repair protein MutL becomes limiting during stationary-phase mutation Genes & Dev., September 15, 1997; 11(18): 2426 - 2437. [Abstract] [Full Text] [PDF]

				S. Rangarajan, G. Gudmundsson, Z. Qiu, P. L. Foster, and M. F. Goodman Escherichia coli DNA polymerase II catalyzes chromosomal and episomal DNA synthesis in vivo PNAS, February 4, 1997; 94(3): 946 - 951. [Abstract] [Full Text] [PDF]

				P. Foster and J. Trimarchi Adaptive reversion of a frameshift mutation in Escherichia coli by simple base deletions in homopolymeric runs Science, July 15, 1994; 265(5170): 407 - 409. [Abstract] [PDF]

				S. T. Fitz-Gibbon, H. Ladner, U.-J. Kim, K. O. Stetter, M. I. Simon, and J. H. Miller Genome sequence of the hyperthermophilic crenarchaeon Pyrobaculum aerophilum PNAS, January 22, 2002; 99(2): 984 - 989. [Abstract] [Full Text] [PDF]