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
. 1976 Jun;73(6):1959–1963. doi: 10.1073/pnas.73.6.1959

Isolation and characterization of conditional lethal mutants of Escherichia coli defective in transcription termination factor rho.

A Das, D Court, S Adhya
PMCID: PMC430427  PMID: 132662

Abstract

Polarity suppressor mutants that are conditional lethal for growth have been isolated in E. coli K12. The mutations map between the ilv and cya loci of the E. coli chromosome. Rho factor isolated from one of these ts mutants does not show transcription termination activity at any temperature tested; however, it is found to be temperature sensitive for its poly(C)-dependent ATPase activity. Unlike the previously known polarity suppressor mutants (suA and psu), the rho mutation suppresses all types of polarity. Other interesting properties of these mutants include ultraviolet sensitivity, recombination deficiency, and decreased ability to lysogenize temperate phages lambda and P1. Our results suggest that rho has an essential function in the growth and normal physiology of cells. The rho(ts) mutant allows the growth of phage lambda defective in the N gene. This result supports the model that N gene product prevents transcription termination by antagonizing rho activity.

Full text

PDF
1959

Images in this article

1961Image
on p.1961

Selected References

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

  1. Adhya S. L., Shapiro J. A. The galactose operon of E. coli K-12. I. Structural and pleiotropic mutations of the operon. Genetics. 1969 Jun;62(2):231–247. doi: 10.1093/genetics/62.2.231. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Adhya S., Gottesman M., De Crombrugghe B. Release of polarity in Escherichia coli by gene N of phage lambda: termination and antitermination of transcription. Proc Natl Acad Sci U S A. 1974 Jun;71(6):2534–2538. doi: 10.1073/pnas.71.6.2534. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Ahmed A. Mechanism of reversion of the gal3 mutation of Escherichia coli. Mol Gen Genet. 1975;136(3):243–253. doi: 10.1007/BF00334019. [DOI] [PubMed] [Google Scholar]
  4. Ahmed A., Scraba D. The nature of the gal3 mutation of Escherichia coli. Mol Gen Genet. 1975;136(3):233–242. doi: 10.1007/BF00334018. [DOI] [PubMed] [Google Scholar]
  5. BECKWITH J. RESTORATION OF OPERON ACTIVITY BY SUPPRESSORS. Biochim Biophys Acta. 1963 Sep 17;76:162–164. [PubMed] [Google Scholar]
  6. BUTTIN G. M'ECANISMES R'EGULATEURS DANS LA BIOSYNTH'ESE DES ENZYMES DU M'ETABOLISME DU GALACTOSE CHEZ ESCHERICHIA COLI K12. II. LE D'ETERMINISME G'EN'ETIQUE DE LA R'EGULATION. J Mol Biol. 1963 Aug;7:183–205. doi: 10.1016/s0022-2836(63)80045-5. [DOI] [PubMed] [Google Scholar]
  7. Brunel F., Davison J. Bacterial mutants able to partly suppress the effect of N mutations in bacteriophage lambda. Mol Gen Genet. 1975;136(2):167–180. doi: 10.1007/BF00272037. [DOI] [PubMed] [Google Scholar]
  8. CAMPBELL A. Sensitive mutants of bacteriophage lambda. Virology. 1961 May;14:22–32. doi: 10.1016/0042-6822(61)90128-3. [DOI] [PubMed] [Google Scholar]
  9. Carter T., Newton A. New polarity suppressors in Escherichia coli: suppression and messenger RNA stability. Proc Natl Acad Sci U S A. 1971 Dec;68(12):2962–2966. doi: 10.1073/pnas.68.12.2962. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. De Crombrugghe B., Adhya S., Gottesman M., Pastan I. Effect of Rho on transcription of bacterial operons. Nat New Biol. 1973 Feb 28;241(113):260–264. doi: 10.1038/newbio241260a0. [DOI] [PubMed] [Google Scholar]
  11. ECHOLS H., REZNICHEK J., ADHYA S. COMPLEMENTATION, RECOMBINATION, AND SUPPRESSION IN GALACTOSE NEGATIVE MUTANTS OF E. COLI. Proc Natl Acad Sci U S A. 1963 Aug;50:286–293. doi: 10.1073/pnas.50.2.286. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. FRANKLIN N. C., LURIA S. E. Transduction by bacteriophage P-1 and the properties of the lac genetic region in E. coli and S. dysenteriae. Virology. 1961 Nov;15:299–311. doi: 10.1016/0042-6822(61)90362-2. [DOI] [PubMed] [Google Scholar]
  13. Fiandt M., Szybalski W., Malamy M. H. Polar mutations in lac, gal and phage lambda consist of a few IS-DNA sequences inserted with either orientation. Mol Gen Genet. 1972;119(3):223–231. doi: 10.1007/BF00333860. [DOI] [PubMed] [Google Scholar]
  14. Gottesman M. E., Yarmolinsky M. B. Integration-negative mutants of bacteriophage lambda. J Mol Biol. 1968 Feb 14;31(3):487–505. doi: 10.1016/0022-2836(68)90423-3. [DOI] [PubMed] [Google Scholar]
  15. Hill C. W., Echols H. Properties of a mutant blocked in inducibility of messenger RNA for the galactose operon of Escherichia coli. J Mol Biol. 1966 Aug;19(1):38–51. doi: 10.1016/s0022-2836(66)80048-7. [DOI] [PubMed] [Google Scholar]
  16. Howard B. H., de Crombrugghe B. ATPase activity required for termination of transcription by the Escherichia coli protein factor rho. J Biol Chem. 1976 Apr 25;251(8):2520–2524. [PubMed] [Google Scholar]
  17. Inoko H., Imai M. Isolation and genetic characterization of the nitA mutants of Escherichia coli affecting the termination factor rho. Mol Gen Genet. 1976 Jan 16;143(2):211–221. doi: 10.1007/BF00266924. [DOI] [PubMed] [Google Scholar]
  18. JACOB F., MONOD J. Genetic regulatory mechanisms in the synthesis of proteins. J Mol Biol. 1961 Jun;3:318–356. doi: 10.1016/s0022-2836(61)80072-7. [DOI] [PubMed] [Google Scholar]
  19. Jordan E., Saedler H., Starlinger P. O0 and strong-polar mutations in the gal operon are insertions. Mol Gen Genet. 1968;102(4):353–363. doi: 10.1007/BF00433726. [DOI] [PubMed] [Google Scholar]
  20. Kourilsky P., Marcaud L., Sheldrick P., Luzzati D., Gros F. Studies of the messenger RNA of bacteriophage lambda, I. Various species synthesized early after induction of the prophage. Proc Natl Acad Sci U S A. 1968 Nov;61(3):1013–1020. doi: 10.1073/pnas.61.3.1013. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Kumar S., Bovre K., Guha A., Hradecna Z., Maher V. R., Szybalski W. Orientation and control of transcription in E. coli phage lambda. Nature. 1969 Mar 1;221(5183):823–825. doi: 10.1038/221823a0. [DOI] [PubMed] [Google Scholar]
  22. 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]
  23. Lieberman R. P., Oishi M. The recBC deoxyribonuclease of Escherichia coli: isolation and characterization of the subunit proteins and reconstitution of the enzyme. Proc Natl Acad Sci U S A. 1974 Dec;71(12):4816–4820. doi: 10.1073/pnas.71.12.4816. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Lowery-Goldhammer C., Richardson J. P. An RNA-dependent nucleoside triphosphate phosphohydrolase (ATPase) associated with rho termination factor. Proc Natl Acad Sci U S A. 1974 May;71(5):2003–2007. doi: 10.1073/pnas.71.5.2003. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Morse D. E., Guertin M. Amber suA mutations which relieve polarity. J Mol Biol. 1972 Feb 14;63(3):605–608. doi: 10.1016/0022-2836(72)90453-6. [DOI] [PubMed] [Google Scholar]
  26. Otsubo E., Lee H. J., Deonier R. C., Davidson N. Electron microscope heteroduplex studies of sequence relations among plasmids of Escherichia coli. VI. Mapping of F14 sequences homologous to phi 80dmetBJF and phi 80dargECBH bacteriophages. J Mol Biol. 1974 Nov 15;89(4):599–618. doi: 10.1016/0022-2836(74)90038-2. [DOI] [PubMed] [Google Scholar]
  27. Ratner D. Evidence that mutations in the suA polarity suppressing gene directly affect termination factor rho. Nature. 1976 Jan 15;259(5539):151–153. doi: 10.1038/259151a0. [DOI] [PubMed] [Google Scholar]
  28. Richardson J. P., Grimley C., Lowery C. Transcription termination factor rho activity is altered in Escherichia coli with suA gene mutations. Proc Natl Acad Sci U S A. 1975 May;72(5):1725–1728. doi: 10.1073/pnas.72.5.1725. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Roberts J. W. Termination factor for RNA synthesis. Nature. 1969 Dec 20;224(5225):1168–1174. doi: 10.1038/2241168a0. [DOI] [PubMed] [Google Scholar]
  30. Rosner J. L. Formation, induction, and curing of bacteriophage P1 lysogens. Virology. 1972 Jun;48(3):679–689. doi: 10.1016/0042-6822(72)90152-3. [DOI] [PubMed] [Google Scholar]
  31. Shapiro J. A., Adhya S. L. The galactose operon of E. coli K-12. II. A deletion analysis of operon structure and polarity. Genetics. 1969 Jun;62(2):249–264. doi: 10.1093/genetics/62.2.249. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Shapiro J. A. Mutations caused by the insertion of genetic material into the galactose operon of Escherichia coli. J Mol Biol. 1969 Feb 28;40(1):93–105. doi: 10.1016/0022-2836(69)90298-8. [DOI] [PubMed] [Google Scholar]
  33. Shimada K., Weisberg R. A., Gottesman M. E. Prophage lambda at unusual chromosomal locations. I. Location of the secondary attachment sites and the properties of the lysogens. J Mol Biol. 1972 Feb 14;63(3):483–503. doi: 10.1016/0022-2836(72)90443-3. [DOI] [PubMed] [Google Scholar]
  34. Shimizu N., Hayashi M. In vitro transcription of the tryptophan operon integrated into a transducing phage genome. J Mol Biol. 1974 Apr 5;84(2):315–335. doi: 10.1016/0022-2836(74)90587-7. [DOI] [PubMed] [Google Scholar]
  35. Shulman M. J., Hallick L. M., Echols H., Signer E. R. Properties of recombination-deficient mutants of bacteriophage lambda. J Mol Biol. 1970 Sep 28;52(3):501–520. doi: 10.1016/0022-2836(70)90416-x. [DOI] [PubMed] [Google Scholar]
  36. Skalka A., Butler B., Echols H. Genetic control of transcription during development of phage gamma. Proc Natl Acad Sci U S A. 1967 Aug;58(2):576–583. doi: 10.1073/pnas.58.2.576. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Weber K., Osborn M. The reliability of molecular weight determinations by dodecyl sulfate-polyacrylamide gel electrophoresis. J Biol Chem. 1969 Aug 25;244(16):4406–4412. [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