Skip to main content
NCBI home page
Proceedings of the National Academy of Sciences of the United States of America logoLink to Proceedings of the National Academy of Sciences of the United States of America
. 1973 Jan;70(1):71–75. doi: 10.1073/pnas.70.1.71

Alteration of Ribosomal Protein S4 by Mutation Linked to Kasugamycin-Resistance in Escherichia coli

Robert A Zimmermann *, Yoshiko Ikeya †, P Frederick Sparling †,§
PMCID: PMC433186  PMID: 4567337

Abstract

An alteration in the chromatographic mobility of 30S ribosomal protein S4 from two kasugamycin-resistant (ksgA) strains of E. coli was observed. The locus determining this alteration frequently cotransduced with ksgA but could be separated from it by transduction and conjugation. Since the structural gene for protein S4 is thought to lie some 30 min from ksgA on the E. coli chromosome, the product of the newly-identified gene may modify protein S4. We propose to designate this gene ramB.

Reconstitution of 30S subunits from RNA and protein components of ksgA ramB and ksgA+ramB+ strains demonstrated that ribosomal resistance to kasugamycin was due to altered 16S RNA and not to altered protein S4. The altered 16S RNA was undermethylated, and a methylating enzyme that acts on 16S RNA from ksgA strains was present in ksgA+ but not in ksgA strains.

Keywords: ribosomal proteins, ribosomal RNA, reconstitution, phosphocellulose chromatography

Full text

PDF
71

Images in this article

73Image
on p.73

Selected References

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

  1. Birge E. A., Kurland C. G. Reversion of a streptomycin-dependent strain of Escherichia coli. Mol Gen Genet. 1970;109(4):356–369. doi: 10.1007/BF00267704. [DOI] [PubMed] [Google Scholar]
  2. Bollen A., Davies J., Ozaki M., Mizushima S. Ribosomal Protein Conferring Sensitivity to the Antibiotic Spectinomycin in Escherichia coli. Science. 1969 Jul 4;165(3888):85–86. doi: 10.1126/science.165.3888.85. [DOI] [PubMed] [Google Scholar]
  3. Cerdá-Olmedo E., Hanawalt P. C., Guerola N. Mutagenesis of the replication point by nitrosoguanidine: map and pattern of replication of the Escherichia coli chromosome. J Mol Biol. 1968 May 14;33(3):705–719. doi: 10.1016/0022-2836(68)90315-x. [DOI] [PubMed] [Google Scholar]
  4. Dekio S. Genetic studies of the ribosomal proteins in Escherichia coli. 7. Mapping of several ribosomal protein components by transduction experiments between different strains of Escherichia coli. Mol Gen Genet. 1971;113(1):20–30. [PubMed] [Google Scholar]
  5. Deusser E., Stöffler G., Wittmann H. G. Ribosomal proteins. XVI. Altered S4 proteins in Escherichia coli revertants from streptomycin dependence to independence. Mol Gen Genet. 1970;109(4):298–302. doi: 10.1007/BF00267699. [DOI] [PubMed] [Google Scholar]
  6. Dzionara M., Kaltschmidt E., Wittmann H. G. Ribosomal proteins. 8. Molecular weights of isolated ribosomal proteins of Escherichia coli. Proc Natl Acad Sci U S A. 1970 Dec;67(4):1909–1913. doi: 10.1073/pnas.67.4.1909. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Fellner P. Nucleotide sequences from specific areas of the 16S and 23S ribosomal RNAs of E. coli. Eur J Biochem. 1969 Nov;11(1):12–27. doi: 10.1111/j.1432-1033.1969.tb00733.x. [DOI] [PubMed] [Google Scholar]
  8. Folk W. R., Berg P. Duplication of the structural gene for glycyl-transfer RNA synthetase in Escherichia coli. J Mol Biol. 1971 Jun 14;58(2):595–610. doi: 10.1016/0022-2836(71)90374-3. [DOI] [PubMed] [Google Scholar]
  9. Funatsu G., Puls W., Schiltz E., Reinbolt J., Wittmann H. G. Ribosomal proteins. XXXI. Comparative studies on altered proteins S4 of six Escherichia coli revertants from streptomycin dependence. Mol Gen Genet. 1972;115(2):131–139. doi: 10.1007/BF00277293. [DOI] [PubMed] [Google Scholar]
  10. Funatsu G., Schiltz E., Wittmann H. G. Ribosomal proteins. XXVII. Localization of the amino acid exchanges in protein S5 from two Escherichia coli mutants resistant to spectinomycin. Mol Gen Genet. 1972;114(2):106–111. doi: 10.1007/BF00332781. [DOI] [PubMed] [Google Scholar]
  11. Hardy S. J., Kurland C. G., Voynow P., Mora G. The ribosomal proteins of Escherichia coli. I. Purification of the 30S ribosomal proteins. Biochemistry. 1969 Jul;8(7):2897–2905. doi: 10.1021/bi00835a031. [DOI] [PubMed] [Google Scholar]
  12. Helser T. L., Davies J. E., Dahlberg J. E. Change in methylation of 16S ribosomal RNA associated with mutation to kasugamycin resistance in Escherichia coli. Nat New Biol. 1971 Sep 1;233(35):12–14. doi: 10.1038/newbio233012a0. [DOI] [PubMed] [Google Scholar]
  13. Helser T. L., Davies J. E., Dahlberg J. E. Mechanism of kasugamycin resistance in Escherichia coli. Nat New Biol. 1972 Jan 5;235(53):6–9. doi: 10.1038/newbio235006a0. [DOI] [PubMed] [Google Scholar]
  14. Kreider G., Brownstein B. L. A mutation suppressing streptomycin dependence. II. An altered protein on the 30 s ribosomal subunit. J Mol Biol. 1971 Oct 14;61(1):135–142. doi: 10.1016/0022-2836(71)90211-7. [DOI] [PubMed] [Google Scholar]
  15. LENNOX E. S. Transduction of linked genetic characters of the host by bacteriophage P1. Virology. 1955 Jul;1(2):190–206. doi: 10.1016/0042-6822(55)90016-7. [DOI] [PubMed] [Google Scholar]
  16. Leibowitz M. J., Soffer R. L. Modification of a specific ribosomal protein catalyzed by leucyl, phenylalanyl-tRNA: protein transferase. Proc Natl Acad Sci U S A. 1971 Aug;68(8):1866–1869. doi: 10.1073/pnas.68.8.1866. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Lowry C. V., Dahlberg J. E. Structural differences between the 16S ribosomal RNA of E. coli and its precursor. Nat New Biol. 1971 Jul 14;232(28):52–54. doi: 10.1038/newbio232052a0. [DOI] [PubMed] [Google Scholar]
  18. Mizushima S., Nomura M. Assembly mapping of 30S ribosomal proteins from E. coli. Nature. 1970 Jun 27;226(5252):1214–1214. doi: 10.1038/2261214a0. [DOI] [PubMed] [Google Scholar]
  19. Moore P. B., Traut R. R., Noller H., Pearson P., Delius H. Ribosomal proteins of Escherichia coli. II. Proteins from the 30 s subunit. J Mol Biol. 1968 Feb 14;31(3):441–461. doi: 10.1016/0022-2836(68)90420-8. [DOI] [PubMed] [Google Scholar]
  20. Nomura M., Mizushima S., Ozaki M., Traub P., Lowry C. V. Structure and function of ribosomes and their molecular components. Cold Spring Harb Symp Quant Biol. 1969;34:49–61. doi: 10.1101/sqb.1969.034.01.009. [DOI] [PubMed] [Google Scholar]
  21. 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]
  22. Paik W. K., Kim S. Protein methylation. Science. 1971 Oct 8;174(4005):114–119. doi: 10.1126/science.174.4005.114. [DOI] [PubMed] [Google Scholar]
  23. 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]
  24. Sparling P. F. Kasugamycin resistance: 30S ribosomal mutation with an unusual location on the Escherichia coli chromosome. Science. 1970 Jan 2;167(3914):56–58. doi: 10.1126/science.167.3914.56. [DOI] [PubMed] [Google Scholar]
  25. Sparling P. F., Modolell J., Takeda Y., Davis B. D. Ribosomes from Escherichia coli merodiplods heterozygous for resistance to streptomycin and to spectinomycin. J Mol Biol. 1968 Nov 14;37(3):407–421. doi: 10.1016/0022-2836(68)90111-3. [DOI] [PubMed] [Google Scholar]
  26. Staehelin T., Meselson M. Determination of streptomycin sensitivity by a subunit of the 30S ribosome of Escherichia coli. J Mol Biol. 1966 Aug;19(1):207–210. doi: 10.1016/s0022-2836(66)80062-1. [DOI] [PubMed] [Google Scholar]
  27. Stöffler G., Daya L., Rak K. H., Garrett R. A. Ribosomal proteins. XXVI. The number of specific protein binding sites on 16 s and 23 s RNA of Escherichia coli. J Mol Biol. 1971 Dec 14;62(2):411–414. doi: 10.1016/0022-2836(71)90437-2. [DOI] [PubMed] [Google Scholar]
  28. Traub P., Nomura M. Structure and function of Escherichia coli ribosomes. VI. Mechanism of assembly of 30 s ribosomes studied in vitro. J Mol Biol. 1969 Mar 28;40(3):391–413. doi: 10.1016/0022-2836(69)90161-2. [DOI] [PubMed] [Google Scholar]
  29. Walton G. M., Gill G. N., Abrass I. B., Garren L. D. Phosphorylation of ribosome-associated protein by an adenosine 3':5'-cyclic monophosphate-dependent protein kinase: location of the microsomal receptor and protein kinase. Proc Natl Acad Sci U S A. 1971 May;68(5):880–884. doi: 10.1073/pnas.68.5.880. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Wittmann H. G., Stöfflet G., Hindennach I., Kurland C. G., Birge E. A., Randall-Hazelbauer L., Nomura M., Kaltschmidt E., Mizushima S., Traut R. R. Correlation of 30S ribosomal proteins of Escherichia coli isolated in different laboratories. Mol Gen Genet. 1971;111(4):327–333. doi: 10.1007/BF00569784. [DOI] [PubMed] [Google Scholar]
  31. 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]

Articles from Proceedings of the National Academy of Sciences of the United States of America are provided here courtesy of National Academy of Sciences

RESOURCES