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. 1972 May;69(5):1192–1195. doi: 10.1073/pnas.69.5.1192

Low Activity of β-Galactosidase in Frameshift Mutants of Escherichia coli

John F Atkins 1,2,*, Dirk Elseviers 1,2, Luigi Gorini 1,2,†
PMCID: PMC426661  PMID: 4556457

Abstract

16 lac frameshift mutants induced by an acridine derivative, ICR-191D, in E. coli are leaky for β-galactosidase activity. Activities of all mutants differ from each other and from the wild type in their stability to thermal denaturation. The leakiness is under ribosomal control, since it is strongly reduced by strA restrictive mutations and is restored by ram mutations that reverse restriction. Addition of streptomycin during growth has an effect similar to the presence of the ram mutation. These ribosomal alterations do not modify the thermal stability of the enzyme.

It is suggested that the leakiness is due to an infrequent 2- or 4-base reading close to the frameshift mutation site. The possibility that not only the ribosome, but also the reading context in the messenger, plays a role in securing code fidelity is discussed.

Keywords: lac operon, ribosomal ambiguity, translational leakiness, acridine mutagenesis

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. BECKWITH J. RESTORATION OF OPERON ACTIVITY BY SUPPRESSORS. Biochim Biophys Acta. 1963 Sep 17;76:162–164. [PubMed] [Google Scholar]
  2. Biswas D. K., Gorini L. Restriction, de-restriction and mistranslation in missense suppression. Ribosomal discrimination of transfer RNA's. J Mol Biol. 1972 Feb 28;64(1):119–134. doi: 10.1016/0022-2836(72)90324-5. [DOI] [PubMed] [Google Scholar]
  3. CRICK F. H., BARNETT L., BRENNER S., WATTS-TOBIN R. J. General nature of the genetic code for proteins. Nature. 1961 Dec 30;192:1227–1232. doi: 10.1038/1921227a0. [DOI] [PubMed] [Google Scholar]
  4. DAVIS B. D., MINGIOLI E. S. Mutants of Escherichia coli requiring methionine or vitamin B12. J Bacteriol. 1950 Jul;60(1):17–28. doi: 10.1128/jb.60.1.17-28.1950. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Epstein W. Transposition of the lac region of Escherichia coli. IV. Escape from repression in bacteriophage-carried lac genes. J Mol Biol. 1967 Dec 28;30(3):529–543. doi: 10.1016/0022-2836(67)90366-x. [DOI] [PubMed] [Google Scholar]
  6. Gorini L. Ribosomal discrimination of tRNAs. Nat New Biol. 1971 Dec 29;234(52):261–264. doi: 10.1038/newbio234261a0. [DOI] [PubMed] [Google Scholar]
  7. Gorini L. The contrasting role of strA and ram gene products in ribosomal functioning. Cold Spring Harb Symp Quant Biol. 1969;34:101–109. doi: 10.1101/sqb.1969.034.01.016. [DOI] [PubMed] [Google Scholar]
  8. Grodzicker T., Zipser D. A mutation which creates a new site for the re-initiation of polypeptide synthesis in the z gene of the lac operon of Escherichia coli. J Mol Biol. 1968 Dec;38(3):305–314. doi: 10.1016/0022-2836(68)90388-4. [DOI] [PubMed] [Google Scholar]
  9. LOWRY O. H., ROSEBROUGH N. J., FARR A. L., RANDALL R. J. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed] [Google Scholar]
  10. Michels C. A., Zipser D. Mapping of polypeptide reinitiation sites within the beta-galactosidase structural gene. J Mol Biol. 1969 May 14;41(3):341–347. doi: 10.1016/0022-2836(69)90280-0. [DOI] [PubMed] [Google Scholar]
  11. Newton A. Isolation and characterization of frameshift mutations in the lac operon. J Mol Biol. 1970 May 14;49(3):589–601. doi: 10.1016/0022-2836(70)90283-4. [DOI] [PubMed] [Google Scholar]
  12. Newton A. Re-initiation of polypeptide synthesis and polarity in the lac operon of Escherichia coli. J Mol Biol. 1969 May 14;41(3):329–339. doi: 10.1016/0022-2836(69)90279-4. [DOI] [PubMed] [Google Scholar]
  13. Ozaki M., Mizushima S., Nomura M. Identification and functional characterization of the protein controlled by the streptomycin-resistant locus in E. coli. Nature. 1969 Apr 26;222(5191):333–339. doi: 10.1038/222333a0. [DOI] [PubMed] [Google Scholar]
  14. Riddle D. L., Roth J. R. Suppressors of frameshift mutations in Salmonella typhimurium. J Mol Biol. 1970 Nov 28;54(1):131–144. doi: 10.1016/0022-2836(70)90451-1. [DOI] [PubMed] [Google Scholar]
  15. Riyasaty S., Atkins J. F. External suppression of a frameshift mutant in salmonella. J Mol Biol. 1968 Jun 28;34(3):541–557. doi: 10.1016/0022-2836(68)90179-4. [DOI] [PubMed] [Google Scholar]
  16. Rosset R., Gorini L. A ribosomal ambiguity mutation. J Mol Biol. 1969 Jan 14;39(1):95–112. doi: 10.1016/0022-2836(69)90336-2. [DOI] [PubMed] [Google Scholar]
  17. Salser W. The influence of the reading context upon the suppression of nonsense codons. Mol Gen Genet. 1969 Oct 13;105(2):125–130. doi: 10.1007/BF00445682. [DOI] [PubMed] [Google Scholar]
  18. Streisinger G., Okada Y., Emrich J., Newton J., Tsugita A., Terzaghi E., Inouye M. Frameshift mutations and the genetic code. This paper is dedicated to Professor Theodosius Dobzhansky on the occasion of his 66th birthday. Cold Spring Harb Symp Quant Biol. 1966;31:77–84. doi: 10.1101/sqb.1966.031.01.014. [DOI] [PubMed] [Google Scholar]
  19. Strigini P., Gorini L. Ribosomal mutations affecting efficiency of amber suppression. J Mol Biol. 1970 Feb 14;47(3):517–530. doi: 10.1016/0022-2836(70)90319-0. [DOI] [PubMed] [Google Scholar]
  20. Tanemura S., Yourno J. Frameshift revertant of Salmonella typhimurium producing histidinol dehydrogenase with a sequence of four extra amino acid residues. J Mol Biol. 1969 Dec 28;46(3):459–466. doi: 10.1016/0022-2836(69)90189-2. [DOI] [PubMed] [Google Scholar]
  21. Terzaghi E., Okada Y., Streisinger G., Emrich J., Inouye M., Tsugita A. Change of a sequence of amino acids in phage T4 lysozyme by acridine-induced mutations. Proc Natl Acad Sci U S A. 1966 Aug;56(2):500–507. doi: 10.1073/pnas.56.2.500. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Ullmann A., Perrin D., Jacob F., Monod J. Identification par complémentation in vitro et purification d'un segment peptidique de la beta-galatosidase d'escherichia coli. J Mol Biol. 1965 Jul;12(3):918–923. doi: 10.1016/s0022-2836(65)80338-2. [DOI] [PubMed] [Google Scholar]
  23. Yourno J., Heath S. Nature of the hisD3018 frameshift mutation in Salmonella typhimurium. J Bacteriol. 1969 Oct;100(1):460–468. doi: 10.1128/jb.100.1.460-468.1969. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Yourno J., Kohno T. Externally suppressible proline quadruplet ccc U. Science. 1972 Feb 11;175(4022):650–652. doi: 10.1126/science.175.4022.650. [DOI] [PubMed] [Google Scholar]
  25. Yourno J., Tanemura S. Restoration of in-phase translation by an unlinked suppressor of a frameshift mutation in Salmonella typhimurium. Nature. 1970 Jan 31;225(5231):422–426. doi: 10.1038/225422a0. [DOI] [PubMed] [Google Scholar]
  26. Zimmermann R. A., Garvin R. T., Gorini L. Alteration of a 30S ribosomal protein accompanying the ram mutation in Escherichia coli. Proc Natl Acad Sci U S A. 1971 Sep;68(9):2263–2267. doi: 10.1073/pnas.68.9.2263. [DOI] [PMC free article] [PubMed] [Google Scholar]

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