Skip to main content
Journal of Bacteriology logoLink to Journal of Bacteriology
. 1984 May;158(2):523–529. doi: 10.1128/jb.158.2.523-529.1984

Genetic analysis of ColN plasmid determinants for colicin production, release, and immunity.

A P Pugsley
PMCID: PMC215460  PMID: 6327609

Abstract

Colicin N was identified as the 39,000-molecular-weight protein encoded by the 4,900-base-pair, multiple copy number, amplifiable plasmid ColN -284. Its production was controlled by the SOS regulatory circuit and by catabolite repression. Colicin accumulated intracellularly to ca. 10(6) molecules per cell after growth for 2 to 3 h in medium containing 0.5 microgram of mitomycin C per ml and was then released as the cells underwent partial lysis. Strains carrying pColN -284 and its derivatives exhibited low-level immunity to colicin N and were fully sensitive to all other colicins tested. Regions of the plasmid responsible for colicin N activity (cna), for mitomycin-induced lysis ( cnl ), and for colicin N immunity ( cni ) were localized and characterized by cloning, transposon Tn5 and hydroxylamine mutagenesis, and restriction endonuclease deletion and mapping analysis. The results are discussed in terms of both the organization of the cna, cnl , and cni genes and the respective role of cnl expression and colicin N production in mitomycin sensitivity, colicin export, and induced partial lysis of ColN + cells.

Full text

PDF
523

Images in this article

Selected References

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

  1. Adhya S., Garges S. How cyclic AMP and its receptor protein act in Escherichia coli. Cell. 1982 Jun;29(2):287–289. doi: 10.1016/0092-8674(82)90145-3. [DOI] [PubMed] [Google Scholar]
  2. Dankert J. R., Uratani Y., Grabau C., Cramer W. A., Hermodson M. On a domain structure of colicin E1. A COOH-terminal peptide fragment active in membrane depolarization. J Biol Chem. 1982 Apr 10;257(7):3857–3863. [PubMed] [Google Scholar]
  3. Davies J. K., Reeves P. Genetics of resistance to colicins in Escherichia coli K-12: cross-resistance among colicins of group A. J Bacteriol. 1975 Jul;123(1):102–117. doi: 10.1128/jb.123.1.102-117.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. HAMON Y., PERON Y. A PROPOS DE QUELQUES NOUVEAUX TYPES DE COLICINES THERMOSTABLES. C R Hebd Seances Acad Sci. 1964 Mar 16;258:3121–3124. [PubMed] [Google Scholar]
  5. Hakkaart M. J., Veltkamp E., Nijkamp H. J. Protein H encoded by plasmid Clo DF13 involved in lysis of the bacterial host. II. Functions and regulation of synthesis of the gene H product. Mol Gen Genet. 1981;183(2):326–332. doi: 10.1007/BF00270636. [DOI] [PubMed] [Google Scholar]
  6. Jorgensen R. A., Rothstein S. J., Reznikoff W. S. A restriction enzyme cleavage map of Tn5 and location of a region encoding neomycin resistance. Mol Gen Genet. 1979;177(1):65–72. doi: 10.1007/BF00267254. [DOI] [PubMed] [Google Scholar]
  7. Little J. W., Mount D. W. The SOS regulatory system of Escherichia coli. Cell. 1982 May;29(1):11–22. doi: 10.1016/0092-8674(82)90085-x. [DOI] [PubMed] [Google Scholar]
  8. Mock M., Pugsley A. P. The BtuB group col plasmids and homology between the colicins they encode. J Bacteriol. 1982 Jun;150(3):1069–1076. doi: 10.1128/jb.150.3.1069-1076.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Pugsley A. P., Rosenbusch J. P. Release of colicin E2 from Escherichia coli. J Bacteriol. 1981 Jul;147(1):186–192. doi: 10.1128/jb.147.1.186-192.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Pugsley A. P., Schnaitman C. A. Identification of three genes controlling production of new outer membrane pore proteins in Escherichia coli K-12. J Bacteriol. 1978 Sep;135(3):1118–1129. doi: 10.1128/jb.135.3.1118-1129.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Pugsley A. P., Schwartz M. A genetic approach to the study of mitomycin-induced lysis of Escherichia coli K-12 strains which produce colicin E2. Mol Gen Genet. 1983;190(3):366–372. doi: 10.1007/BF00331060. [DOI] [PubMed] [Google Scholar]
  12. Pugsley A. P. Transcription regulation of colicin Ib synthesis. Mol Gen Genet. 1981;183(3):522–527. doi: 10.1007/BF00268775. [DOI] [PubMed] [Google Scholar]
  13. 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]
  14. Sabik J. F., Suit J. L., Luria S. E. cea-kil operon of the ColE1 plasmid. J Bacteriol. 1983 Mar;153(3):1479–1485. doi: 10.1128/jb.153.3.1479-1485.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Suit J. L., Fan M. L., Sabik J. F., Labarre R., Luria S. E. Alternative forms of lethality in mitomycin C-induced bacteria carrying ColE1 plasmids. Proc Natl Acad Sci U S A. 1983 Jan;80(2):579–583. doi: 10.1073/pnas.80.2.579. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Sutcliffe J. G. Complete nucleotide sequence of the Escherichia coli plasmid pBR322. Cold Spring Harb Symp Quant Biol. 1979;43(Pt 1):77–90. doi: 10.1101/sqb.1979.043.01.013. [DOI] [PubMed] [Google Scholar]
  17. Völker T. A., Showe M. K. Induction of mutations in specific genes of bacteriophage T4 using cloned restriction fragments and marker rescue. Mol Gen Genet. 1980 Feb;177(3):447–452. doi: 10.1007/BF00271483. [DOI] [PubMed] [Google Scholar]
  18. Watson R. J., Visentin L. P. Cloning of the ColE3-CA38 colicin and immunity genes and identification of a plasmid region which enhances colicin production. Gene. 1982 Sep;19(2):191–200. doi: 10.1016/0378-1119(82)90006-3. [DOI] [PubMed] [Google Scholar]
  19. Watson R., Rowsome W., Tsao J., Visentin L. P. Identification and characterization of Col plasmids from classical colicin E-producing strains. J Bacteriol. 1981 Aug;147(2):569–577. doi: 10.1128/jb.147.2.569-577.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]

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

RESOURCES