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
. 1989 Jun;86(11):4002–4006. doi: 10.1073/pnas.86.11.4002

Translational repression in bacteriophage f1: characterization of the gene V protein target on the gene II mRNA.

B Michel 1, N D Zinder 1
PMCID: PMC287376  PMID: 2657734

Abstract

Previous studies have shown that the single-stranded DNA binding protein of bacteriophage f1 (gene V protein) represses the translation of the mRNA of the phage-encoded replication protein (gene II protein). We have characterized phage mutations in the repressor and in its target. Using a gene II-lacZ translational fusion, we have defined a 16-nucleotide-long region in the gene II mRNA sequence that is required in vivo for repression by the gene V protein. We have shown that in vitro the binding affinity of the gene V protein is at least 10-fold higher to an RNA carrying this sequence than to an RNA lacking it. We propose that this sequence constitutes the gene II mRNA operator.

Full text

PDF
4002

Images in this article

4003Image
on p.4003
4005Image
on p.4005
4005Image
on p.4005

Selected References

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

  1. Alberts B., Frey L., Delius H. Isolation and characterization of gene 5 protein of filamentous bacterial viruses. J Mol Biol. 1972 Jul 14;68(1):139–152. doi: 10.1016/0022-2836(72)90269-0. [DOI] [PubMed] [Google Scholar]
  2. Alma N. C., Harmsen B. J., de Jong E. A., Ven J., Hilbers C. W. Fluorescence studies of the complex formation between the gene 5 protein of bacteriophage M13 and polynucleotides. J Mol Biol. 1983 Jan 5;163(1):47–62. doi: 10.1016/0022-2836(83)90029-3. [DOI] [PubMed] [Google Scholar]
  3. Alma N. C., Harmsen B. J., van Boom J. H., van der Marel G., Hilbers C. W. 1H NMR studies of the binding of bacteriophage-M13-encoded gene-5 protein to oligo(deoxyadenylic acid)s of varying length. Eur J Biochem. 1982 Feb;122(2):319–326. doi: 10.1111/j.1432-1033.1982.tb05883.x. [DOI] [PubMed] [Google Scholar]
  4. Birnboim H. C., Doly J. A rapid alkaline extraction procedure for screening recombinant plasmid DNA. Nucleic Acids Res. 1979 Nov 24;7(6):1513–1523. doi: 10.1093/nar/7.6.1513. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Bulsink H., Harmsen B. J., Hilbers C. W. Specificity of the binding of bacteriophage M13 encoded gene-5 protein to DNA and RNA studied by means of fluorescence titrations. J Biomol Struct Dyn. 1985 Oct;3(2):227–247. doi: 10.1080/07391102.1985.10508413. [DOI] [PubMed] [Google Scholar]
  6. Bulsink H., Wijnaendts van Resandt R. W., Harmsen B. J., Hilbers C. W. Different DNA-binding modes and cooperativities for bacteriophage M13 gene-5 protein revealed by means of fluorescence depolarisation studies. Eur J Biochem. 1986 Jun 2;157(2):329–334. doi: 10.1111/j.1432-1033.1986.tb09672.x. [DOI] [PubMed] [Google Scholar]
  7. 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]
  8. Deckman I. C., Draper D. E. S4-alpha mRNA translation regulation complex. II. Secondary structures of the RNA regulatory site in the presence and absence of S4. J Mol Biol. 1987 Jul 20;196(2):323–332. doi: 10.1016/0022-2836(87)90693-0. [DOI] [PubMed] [Google Scholar]
  9. Dotto G. P., Horiuchi K., Zinder N. D. The functional origin of bacteriophage f1 DNA replication. Its signals and domains. J Mol Biol. 1984 Feb 5;172(4):507–521. doi: 10.1016/s0022-2836(84)80020-0. [DOI] [PubMed] [Google Scholar]
  10. Dotto G. P., Zinder N. D. Increased intracellular concentration of an initiator protein markedly reduces the minimal sequence required for initiation of DNA synthesis. Proc Natl Acad Sci U S A. 1984 Mar;81(5):1336–1340. doi: 10.1073/pnas.81.5.1336. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Dotto G. P., Zinder N. D. Reduction of the minimal sequence for initiation of DNA synthesis by qualitative or quantitative changes of an initiator protein. Nature. 1984 Sep 20;311(5983):279–280. doi: 10.1038/311279a0. [DOI] [PubMed] [Google Scholar]
  12. Dreyfus M. What constitutes the signal for the initiation of protein synthesis on Escherichia coli mRNAs? J Mol Biol. 1988 Nov 5;204(1):79–94. doi: 10.1016/0022-2836(88)90601-8. [DOI] [PubMed] [Google Scholar]
  13. Dunker A. K. Analysis of protein-nucleic acid filter binding using the Poisson distribution: a method to estimate co-operativity. FEBS Lett. 1975 Apr 1;52(2):323–326. doi: 10.1016/0014-5793(75)80835-0. [DOI] [PubMed] [Google Scholar]
  14. Fulford W., Model P. Bacteriophage f1 DNA replication genes. II. The roles of gene V protein and gene II protein in complementary strand synthesis. J Mol Biol. 1988 Sep 5;203(1):39–48. doi: 10.1016/0022-2836(88)90089-7. [DOI] [PubMed] [Google Scholar]
  15. Fulford W., Model P. Specificity of translational regulation by two DNA-binding proteins. J Mol Biol. 1984 Feb 25;173(2):211–226. doi: 10.1016/0022-2836(84)90190-6. [DOI] [PubMed] [Google Scholar]
  16. Gold L. Posttranscriptional regulatory mechanisms in Escherichia coli. Annu Rev Biochem. 1988;57:199–233. doi: 10.1146/annurev.bi.57.070188.001215. [DOI] [PubMed] [Google Scholar]
  17. Greenstein D., Horiuchi K. Interaction between the replication origin and the initiator protein of the filamentous phage f1. Binding occurs in two steps. J Mol Biol. 1987 Sep 20;197(2):157–174. doi: 10.1016/0022-2836(87)90115-x. [DOI] [PubMed] [Google Scholar]
  18. Greenstein D., Zinder N. D., Horiuchi K. Integration host factor interacts with the DNA replication enhancer of filamentous phage f1. Proc Natl Acad Sci U S A. 1988 Sep;85(17):6262–6266. doi: 10.1073/pnas.85.17.6262. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Hill D. F., Petersen G. B. Nucleotide sequence of bacteriophage f1 DNA. J Virol. 1982 Oct;44(1):32–46. doi: 10.1128/jvi.44.1.32-46.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Lyons L. B., Zinder N. D. The genetic map of the filamentous bacteriophage f1. Virology. 1972 Jul;49(1):45–60. doi: 10.1016/s0042-6822(72)80006-0. [DOI] [PubMed] [Google Scholar]
  21. Mazur B. J., Zinder N. D. The role of gene V protein in f1 single-strand synthesis. Virology. 1975 Dec;68(2):490–502. doi: 10.1016/0042-6822(75)90289-5. [DOI] [PubMed] [Google Scholar]
  22. McPheeters D. S., Stormo G. D., Gold L. Autogenous regulatory site on the bacteriophage T4 gene 32 messenger RNA. J Mol Biol. 1988 Jun 5;201(3):517–535. doi: 10.1016/0022-2836(88)90634-1. [DOI] [PubMed] [Google Scholar]
  23. Meyer T. F., Geider K. Enzymatic synthesis of bacteriophage fd viral DNA. Nature. 1982 Apr 29;296(5860):828–832. doi: 10.1038/296828a0. [DOI] [PubMed] [Google Scholar]
  24. Meyer T. F., Geider K., Kurz C., Schaller H. Cleavage site of bacteriophage fd gene II-protein in the origin of viral strand replication. Nature. 1979 Mar 22;278(5702):365–367. doi: 10.1038/278365a0. [DOI] [PubMed] [Google Scholar]
  25. Milligan J. F., Groebe D. R., Witherell G. W., Uhlenbeck O. C. Oligoribonucleotide synthesis using T7 RNA polymerase and synthetic DNA templates. Nucleic Acids Res. 1987 Nov 11;15(21):8783–8798. doi: 10.1093/nar/15.21.8783. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Model P., McGill C., Mazur B., Fulford W. D. The replication of bacteriophage f1: gene V protein regulates the synthesis of gene II protein. Cell. 1982 Jun;29(2):329–335. doi: 10.1016/0092-8674(82)90149-0. [DOI] [PubMed] [Google Scholar]
  27. Raleigh E. A., Murray N. E., Revel H., Blumenthal R. M., Westaway D., Reith A. D., Rigby P. W., Elhai J., Hanahan D. McrA and McrB restriction phenotypes of some E. coli strains and implications for gene cloning. Nucleic Acids Res. 1988 Feb 25;16(4):1563–1575. doi: 10.1093/nar/16.4.1563. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Romaniuk P. J., Lowary P., Wu H. N., Stormo G., Uhlenbeck O. C. RNA binding site of R17 coat protein. Biochemistry. 1987 Mar 24;26(6):1563–1568. doi: 10.1021/bi00380a011. [DOI] [PubMed] [Google Scholar]
  29. Rose R. E. The nucleotide sequence of pACYC184. Nucleic Acids Res. 1988 Jan 11;16(1):355–355. doi: 10.1093/nar/16.1.355. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Russel M., Kidd S., Kelley M. R. An improved filamentous helper phage for generating single-stranded plasmid DNA. Gene. 1986;45(3):333–338. doi: 10.1016/0378-1119(86)90032-6. [DOI] [PubMed] [Google Scholar]
  31. Russel M., Model P. The role of thioredoxin in filamentous phage assembly. Construction, isolation, and characterization of mutant thioredoxins. J Biol Chem. 1986 Nov 15;261(32):14997–15005. [PubMed] [Google Scholar]
  32. Sanger F., Nicklen S., Coulson A. R. DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci U S A. 1977 Dec;74(12):5463–5467. doi: 10.1073/pnas.74.12.5463. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Scholtissek S., Grosse F. A cloning cartridge of lambda t(o) terminator. Nucleic Acids Res. 1987 Apr 10;15(7):3185–3185. doi: 10.1093/nar/15.7.3185. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Shapira S. K., Chou J., Richaud F. V., Casadaban M. J. New versatile plasmid vectors for expression of hybrid proteins coded by a cloned gene fused to lacZ gene sequences encoding an enzymatically active carboxy-terminal portion of beta-galactosidase. Gene. 1983 Nov;25(1):71–82. doi: 10.1016/0378-1119(83)90169-5. [DOI] [PubMed] [Google Scholar]
  35. Silhavy T. J., Beckwith J. R. Uses of lac fusions for the study of biological problems. Microbiol Rev. 1985 Dec;49(4):398–418. doi: 10.1128/mr.49.4.398-418.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Steitz J. A. Polypeptide chain initiation: nucleotide sequences of the three ribosomal binding sites in bacteriophage R17 RNA. Nature. 1969 Dec 6;224(5223):957–964. doi: 10.1038/224957a0. [DOI] [PubMed] [Google Scholar]
  37. Vandeyar M. A., Weiner M. P., Hutton C. J., Batt C. A. A simple and rapid method for the selection of oligodeoxynucleotide-directed mutants. Gene. 1988 May 15;65(1):129–133. doi: 10.1016/0378-1119(88)90425-8. [DOI] [PubMed] [Google Scholar]
  38. Yen T. S., Webster R. E. Translational control of bacteriophage f1 gene II and gene X proteins by gene V protein. Cell. 1982 Jun;29(2):337–345. doi: 10.1016/0092-8674(82)90150-7. [DOI] [PubMed] [Google Scholar]
  39. Zinder N. D., Boeke J. D. The filamentous phage (Ff) as vectors for recombinant DNA--a review. Gene. 1982 Jul-Aug;19(1):1–10. doi: 10.1016/0378-1119(82)90183-4. [DOI] [PubMed] [Google Scholar]
  40. von Hippel P. H., Kowalczykowski S. C., Lonberg N., Newport J. W., Paul L. S., Stormo G. D., Gold L. Autoregulation of gene expression. Quantitative evaluation of the expression and function of the bacteriophage T4 gene 32 (single-stranded DNA binding) protein system. J Mol Biol. 1982 Dec 25;162(4):795–818. doi: 10.1016/0022-2836(82)90548-4. [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