Skip to main content
PMC 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
. 1981 May;78(5):2937–2941. doi: 10.1073/pnas.78.5.2937

Mutations that affect lamB gene expression at a posttranscriptional level.

M Schwartz, M Roa, M Débarbouillé
PMCID: PMC319474  PMID: 6265927

Abstract

We previously obtained strains of Escherichia coli in which the beginning of gene lacZ, which codes for beta-galactosidase, is replaced by the beginning of gene lamB, which codes for a maltose-inducible outer membrane protein. In some of these strains the induction (with maltose) of lamB-lacZ hybrid protein synthesis was lethal because of membrane damage resulting from an incomplete export of this protein to the outer membrane. We describe here a class of maltose-resistant mutants obtained from one such strain. Mutants in this class fail to produce the lamB-lacZ hybrid protein but retain the ability to express lacY, which is located distal to the hybrid gene. Some of the mutants carry deletions within the hybrid gene. The others carry point mutations which most probably affect the initiation of translation at the beginning of the hybrid gene. One of these is located in the sequence that codes for the presumed ribosome interaction site on the mRNA. Three others, of which two are located in the coding region (sixth codon), are believed to result in an alteration of mRNA secondary structure such that the accessibility of the ribosome interaction site is reduced.

Full text

PDF
2940

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. BUTTIN G., COHEN G. N., MONOD J., RICKENBERG H. V. La galactoside-perméase d'Escherichia coli. Ann Inst Pasteur (Paris) 1956 Dec;91(6):829–857. [PubMed] [Google Scholar]
  3. Baan R. A., Hilbers C. W., Van Charldorp R., Van Leerdam E., Van Knippenberg P. H., Bosch L. High-resolution proton magnetic resonance study of the secondary structure of the 3'-terminal 49-nucleotide fragment of 16S rRNA from Escherichia coli. Proc Natl Acad Sci U S A. 1977 Mar;74(3):1028–1031. doi: 10.1073/pnas.74.3.1028. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Bassford P. J., Jr, Silhavy T. J., Beckwith J. R. Use of gene fusion to study secretion of maltose-binding protein into Escherichia coli periplasm. J Bacteriol. 1979 Jul;139(1):19–31. doi: 10.1128/jb.139.1.19-31.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Blobel G., Dobberstein B. Transfer of proteins across membranes. I. Presence of proteolytically processed and unprocessed nascent immunoglobulin light chains on membrane-bound ribosomes of murine myeloma. J Cell Biol. 1975 Dec;67(3):835–851. doi: 10.1083/jcb.67.3.835. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Dunn J. J., Buzash-Pollert E., Studier F. W. Mutations of bacteriophage T7 that affect initiation of synthesis of the gene 0.3 protein. Proc Natl Acad Sci U S A. 1978 Jun;75(6):2741–2745. doi: 10.1073/pnas.75.6.2741. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Débarbouillé M., Shuman H. A., Silhavy T. J., Schwartz M. Dominant constitutive mutations in malT, the positive regulator gene of the maltose regulon in Escherichia coli. J Mol Biol. 1978 Sep 15;124(2):359–371. doi: 10.1016/0022-2836(78)90304-2. [DOI] [PubMed] [Google Scholar]
  8. Emr S. D., Hedgpeth J., Clément J. M., Silhavy T. J., Hofnung M. Sequence analysis of mutations that prevent export of lambda receptor, an Escherichia coli outer membrane protein. Nature. 1980 May 8;285(5760):82–85. doi: 10.1038/285082a0. [DOI] [PubMed] [Google Scholar]
  9. Emr S. D., Schwartz M., Silhavy T. J. Mutations altering the cellular localization of the phage lambda receptor, an Escherichia coli outer membrane protein. Proc Natl Acad Sci U S A. 1978 Dec;75(12):5802–5806. doi: 10.1073/pnas.75.12.5802. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Emr S. D., Silhavy T. J. Mutations affecting localization of an Escherichia coli outer membrane protein, the bacteriophage lambda receptor. J Mol Biol. 1980 Jul 25;141(1):63–90. doi: 10.1016/s0022-2836(80)80029-5. [DOI] [PubMed] [Google Scholar]
  11. Hedgpeth J., Clement J. M., Marchal C., Perrin D., Hofnung M. DNA sequence encoding the NH2-terminal peptide involved in transport of lambda receptor, an Escherichia coli secretory protein. Proc Natl Acad Sci U S A. 1980 May;77(5):2621–2625. doi: 10.1073/pnas.77.5.2621. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Iserentant D., Fiers W. Secondary structure of mRNA and efficiency of translation initiation. Gene. 1980 Apr;9(1-2):1–12. doi: 10.1016/0378-1119(80)90163-8. [DOI] [PubMed] [Google Scholar]
  13. Maxam A. M., Gilbert W. A new method for sequencing DNA. Proc Natl Acad Sci U S A. 1977 Feb;74(2):560–564. doi: 10.1073/pnas.74.2.560. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. 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]
  15. Moreno F., Fowler A. V., Hall M., Silhavy T. J., Zabin I., Schwartz M. A signal sequence is not sufficient to lead beta-galactosidase out of the cytoplasm. Nature. 1980 Jul 24;286(5771):356–359. doi: 10.1038/286356a0. [DOI] [PubMed] [Google Scholar]
  16. 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]
  17. PRESTIDGE L. S., PARDEE A. B. A SECOND PERMEASE FOR METHYL-THIO-BETA-D-GALACTOSIDE IN ESCHERICHIA COLI. Biochim Biophys Acta. 1965 May 4;100:591–593. doi: 10.1016/0304-4165(65)90029-2. [DOI] [PubMed] [Google Scholar]
  18. Raibaud O., Clément J. M., Hofnung M. Structure of the malB region in Escherichia coli K12. III. Correlation of the genetic map with the restriction map. Mol Gen Genet. 1979 Jul 24;174(3):261–267. doi: 10.1007/BF00267798. [DOI] [PubMed] [Google Scholar]
  19. Raibaud O., Roa M., Braun-Breton C., Schwartz M. Structure of the malB region in Escherichia coli K12. I. Genetic map of the malK-lamB operon. Mol Gen Genet. 1979 Jul 24;174(3):241–248. doi: 10.1007/BF00267796. [DOI] [PubMed] [Google Scholar]
  20. Randall-Hazelbauer L., Schwartz M. Isolation of the bacteriophage lambda receptor from Escherichia coli. J Bacteriol. 1973 Dec;116(3):1436–1446. doi: 10.1128/jb.116.3.1436-1446.1973. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Roa M., Clément J. M. Location of a phage binding region on an outer membrane protein. FEBS Lett. 1980 Nov 17;121(1):127–129. doi: 10.1016/0014-5793(80)81280-4. [DOI] [PubMed] [Google Scholar]
  22. Roa M. Interaction of bacteriophage K10 with its receptor, the lamB protein of Escherichia coli. J Bacteriol. 1979 Nov;140(2):680–686. doi: 10.1128/jb.140.2.680-686.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Scherer G. F., Walkinshaw M. D., Arnott S., Morré D. J. The ribosome binding sites recognized by E. coli ribosomes have regions with signal character in both the leader and protein coding segments. Nucleic Acids Res. 1980 Sep 11;8(17):3895–3907. doi: 10.1093/nar/8.17.3895. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Schwartz M. The adsorption of coliphage lambda to its host: effect of variations in the surface density of receptor and in phage-receptor affinity. J Mol Biol. 1976 May 25;103(3):521–536. doi: 10.1016/0022-2836(76)90215-1. [DOI] [PubMed] [Google Scholar]
  25. Shine J., Dalgarno L. Determinant of cistron specificity in bacterial ribosomes. Nature. 1975 Mar 6;254(5495):34–38. doi: 10.1038/254034a0. [DOI] [PubMed] [Google Scholar]
  26. Silhavy T. J., Shuman H. A., Beckwith J., Schwartz M. Use of gene fusions to study outer membrane protein localization in Escherichia coli. Proc Natl Acad Sci U S A. 1977 Dec;74(12):5411–5415. doi: 10.1073/pnas.74.12.5411. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Sprague K. U., Steitz J. A., Grenley R. M., Stocking C. E. 3' terminal sequences of 16S rRNA do not explain translational specificity differences between E. coli and B. stearothermophilus ribosomes. Nature. 1977 Jun 2;267(5610):462–465. doi: 10.1038/267462a0. [DOI] [PubMed] [Google Scholar]
  28. Steitz J. A., Steege D. A. Characterization of two mRNA-rRNA complexes implicated in the initiation of protein biosynthesis. J Mol Biol. 1977 Aug 25;114(4):545–558. doi: 10.1016/0022-2836(77)90177-2. [DOI] [PubMed] [Google Scholar]
  29. Szmelcman S., Hofnung M. Maltose transport in Escherichia coli K-12: involvement of the bacteriophage lambda receptor. J Bacteriol. 1975 Oct;124(1):112–118. doi: 10.1128/jb.124.1.112-118.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Tinoco I., Jr, Borer P. N., Dengler B., Levin M. D., Uhlenbeck O. C., Crothers D. M., Bralla J. Improved estimation of secondary structure in ribonucleic acids. Nat New Biol. 1973 Nov 14;246(150):40–41. doi: 10.1038/newbio246040a0. [DOI] [PubMed] [Google Scholar]
  31. Wandersman C., Schwartz M., Ferenci T. Escherichia coli mutants impaired in maltodextrin transport. J Bacteriol. 1979 Oct;140(1):1–13. doi: 10.1128/jb.140.1.1-13.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Wandersman C., Schwartz M. Protein Ia and the lamB protein can replace each other in the constitution of an active receptor for the same coliphage. Proc Natl Acad Sci U S A. 1978 Nov;75(11):5636–5639. doi: 10.1073/pnas.75.11.5636. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Welply J. K., Fowler A. V., Beckwith J. R., Zabin I. Positions of early nonsense and deletion mutations in lacZ. J Bacteriol. 1980 May;142(2):732–734. doi: 10.1128/jb.142.2.732-734.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. von Heijne G. Trans-membrane translocation of proteins. A detailed physico-chemical analysis. Eur J Biochem. 1980 Feb;103(3):431–438. doi: 10.1111/j.1432-1033.1980.tb05966.x. [DOI] [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