Skip to main content
PMC home page
Journal of Bacteriology logoLink to Journal of Bacteriology
. 1983 Aug;155(2):854–859. doi: 10.1128/jb.155.2.854-859.1983

Identification of the rodA gene product of Escherichia coli.

N G Stoker, J M Pratt, B G Spratt
PMCID: PMC217759  PMID: 6348029

Abstract

Plasmids that carry the Escherichia coli cell shape gene rodA directed the synthesis of a cytoplasmic membrane protein (Mr, 31,000 [31K protein] ) in minicells, maxicells, and an in vitro-coupled transcription-translation system. The 31K protein was identified as the rodA gene product, because it was not synthesized from the vector plasmids or from a plasmid in which the rodA gene was inactivated by insertion of Tn1000. Furthermore, a purified 1.6-kilobase KpnI-BamHI DNA fragment that contained the intact rodA gene directed the synthesis of only the 31K protein in an in vitro system. The apparent molecular weight of the protein was identical whether synthesized in vivo or in vitro, indicating that the rodA gene product is not made as a preprotein. The direction of transcription of rodA was from the KpnI site towards the BamHI site. The 31K protein was unusual in that it could only be detected when cell membranes were solubilized at low temperature (e.g., 37 degrees C) before sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Apparently the rodA gene product aggregates after being boiled in sodium dodecyl sulfate and fails to enter a polyacrylamide gel.

Full text

PDF
856

Images in this article

856Image
on p.856
857Image
on p.857
857Image
on p.857
858Image
on p.858

Selected References

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

  1. Bachmann B. J., Low K. B. Linkage map of Escherichia coli K-12, edition 6. Microbiol Rev. 1980 Mar;44(1):1–56. doi: 10.1128/mr.44.1.1-56.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Chang A. C., Cohen S. N. Construction and characterization of amplifiable multicopy DNA cloning vehicles derived from the P15A cryptic miniplasmid. J Bacteriol. 1978 Jun;134(3):1141–1156. doi: 10.1128/jb.134.3.1141-1156.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. De Crombrugghe B., Pastan I., Shaw W. V., Rosner J. L. Stimulation by cyclic AMP and ppGpp of chloramphenicol acetyl transferase synthesis. Nat New Biol. 1973 Feb 21;241(112):237–239. doi: 10.1038/newbio241237a0. [DOI] [PubMed] [Google Scholar]
  4. Ehring R., Beyreuther K., Wright J. K., Overath P. In vitro and in vivo products of E. coli lactose permease gene are identical. Nature. 1980 Feb 7;283(5747):537–540. doi: 10.1038/283537a0. [DOI] [PubMed] [Google Scholar]
  5. Guyer M. S. The gamma delta sequence of F is an insertion sequence. J Mol Biol. 1978 Dec 15;126(3):347–365. doi: 10.1016/0022-2836(78)90045-1. [DOI] [PubMed] [Google Scholar]
  6. Herrero E., Jackson M., Bassford P. J., Sinden D., Holland I. B. Insertion of a MalE beta-galactosidase fusion protein into the envelope of Escherichia coli disrupts biogenesis of outer membrane proteins and processing of inner membrane proteins. J Bacteriol. 1982 Oct;152(1):133–139. doi: 10.1128/jb.152.1.133-139.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Higgins C. F., Haag P. D., Nikaido K., Ardeshir F., Garcia G., Ames G. F. Complete nucleotide sequence and identification of membrane components of the histidine transport operon of S. typhimurium. Nature. 1982 Aug 19;298(5876):723–727. doi: 10.1038/298723a0. [DOI] [PubMed] [Google Scholar]
  8. Ishino F., Tamaki S., Spratt B. G., Matsuhashi M. A mecillinam-sensitive peptidoglycan crosslinking reaction in Escherichia coli. Biochem Biophys Res Commun. 1982 Dec 15;109(3):689–696. doi: 10.1016/0006-291x(82)91995-7. [DOI] [PubMed] [Google Scholar]
  9. Iwaya M., Jones C. W., Khorana J., Strominger J. L. Mapping of the mecillinam-resistant, round morphological mutants of Escherichia coli. J Bacteriol. 1978 Jan;133(1):196–202. doi: 10.1128/jb.133.1.196-202.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Larson T. J., Schumacher G., Boos W. Identification of the glpT-encoded sn-glycerol-3-phosphate permease of Escherichia coli, an oligomeric integral membrane protein. J Bacteriol. 1982 Dec;152(3):1008–1021. doi: 10.1128/jb.152.3.1008-1021.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Lilley D. M. In vivo consequences of plasmid topology. Nature. 1981 Jul 23;292(5821):380–382. doi: 10.1038/292380a0. [DOI] [PubMed] [Google Scholar]
  12. Markiewicz Z., Broome-Smith J. K., Schwarz U., Spratt B. G. Spherical E. coli due to elevated levels of D-alanine carboxypeptidase. Nature. 1982 Jun 24;297(5868):702–704. doi: 10.1038/297702a0. [DOI] [PubMed] [Google Scholar]
  13. Matsuzawa H., Hayakawa K., Sato T., Imahori K. Characterization and genetic analysis of a mutant of Escherichia coli K-12 with rounded morphology. J Bacteriol. 1973 Jul;115(1):436–442. doi: 10.1128/jb.115.1.436-442.1973. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Pratt J. M., Boulnois G. J., Darby V., Orr E., Wahle E., Holland I. B. Identification of gene products programmed by restriction endonuclease DNA fragments using an E. coli in vitro system. Nucleic Acids Res. 1981 Sep 25;9(18):4459–4474. doi: 10.1093/nar/9.18.4459. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Pratt J. M., Holland I. B., Spratt B. G. Precursor forms of penicillin-binding proteins 5 and 6 of E. coli cytoplasmic membrane. Nature. 1981 Sep 24;293(5830):307–309. doi: 10.1038/293307a0. [DOI] [PubMed] [Google Scholar]
  16. Reeve J. Use of minicells for bacteriophage-directed polypeptide synthesis. Methods Enzymol. 1979;68:493–503. doi: 10.1016/0076-6879(79)68038-2. [DOI] [PubMed] [Google Scholar]
  17. Sancar A., Hack A. M., Rupp W. D. Simple method for identification of plasmid-coded proteins. J Bacteriol. 1979 Jan;137(1):692–693. doi: 10.1128/jb.137.1.692-693.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Spratt B. G., Boyd A., Stoker N. Defective and plaque-forming lambda transducing bacteriophage carrying penicillin-binding protein-cell shape genes: genetic and physical mapping and identification of gene products from the lip-dacA-rodA-pbpA-leuS region of the Escherichia coli chromosome. J Bacteriol. 1980 Aug;143(2):569–581. doi: 10.1128/jb.143.2.569-581.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Spratt B. G. Distinct penicillin binding proteins involved in the division, elongation, and shape of Escherichia coli K12. Proc Natl Acad Sci U S A. 1975 Aug;72(8):2999–3003. doi: 10.1073/pnas.72.8.2999. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Spratt B. G. Properties of the penicillin-binding proteins of Escherichia coli K12,. Eur J Biochem. 1977 Jan;72(2):341–352. doi: 10.1111/j.1432-1033.1977.tb11258.x. [DOI] [PubMed] [Google Scholar]
  21. Stoker N. G., Broome-Smith J. K., Edelman A., Spratt B. G. Organization and subcloning of the dacA-rodA-pbpA cluster of cell shape genes in Escherichia coli. J Bacteriol. 1983 Aug;155(2):847–853. doi: 10.1128/jb.155.2.847-853.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Stoker N. G., Fairweather N. F., Spratt B. G. Versatile low-copy-number plasmid vectors for cloning in Escherichia coli. Gene. 1982 Jun;18(3):335–341. doi: 10.1016/0378-1119(82)90172-x. [DOI] [PubMed] [Google Scholar]
  23. Tait R. C., Boyer H. W. On the nature of tetracycline resistance controlled by the plasmid pSC101. Cell. 1978 Jan;13(1):73–81. doi: 10.1016/0092-8674(78)90139-3. [DOI] [PubMed] [Google Scholar]
  24. Tamaki S., Matsuzawa H., Matsuhashi M. Cluster of mrdA and mrdB genes responsible for the rod shape and mecillinam sensitivity of Escherichia coli. J Bacteriol. 1980 Jan;141(1):52–57. doi: 10.1128/jb.141.1.52-57.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Teather R. M., Müller-Hill B., Abrutsch U., Aichele G., Overath P. Amplification of the lactose carrier protein in Escherichia coli using a plasmid vector. Mol Gen Genet. 1978 Feb 27;159(3):239–248. doi: 10.1007/BF00268260. [DOI] [PubMed] [Google Scholar]
  26. Westling-Häggström B., Normark S. Genetic and physiological analysis of an envB spherelike mutant of Escherichia coli K-12 and characterization of its transductants. J Bacteriol. 1975 Jul;123(1):75–82. doi: 10.1128/jb.123.1.75-82.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Journal of Bacteriology are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES