Skip to main content
PMC home page
The EMBO Journal logoLink to The EMBO Journal
. 1982;1(2):205–209. doi: 10.1002/j.1460-2075.1982.tb01148.x

Functional expression of individual plasmid-coded RNA bacteriophage MS2 genes

Erik Remaut 1, Peter De Waele 1, Anne Marmenout 1, Patrick Stanssens 1, Walter Fiers 1,*
PMCID: PMC553021  PMID: 16453413

Abstract

The genes of the RNA-containing bacteriophage MS2 were individually inserted into thermoinducible expression plasmids under control of the phage λ PL promoter. Three phage-coded proteins (A-protein, coat protein, and replicase) were expressed at high efficiency. Induced cultures specifically complemented superinfecting amber mutants of phage MS2. Regulatory mechanisms operative during the natural infection cycle of the phage were reproduced by the plasmid expression system.

Keywords: complementation, MS2 proteins, PL promoter

Full text

PDF
205

Images in this article

206Fig. 2.
on p.206

Selected References

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

  1. APPLEYARD R. K., MCGREGOR J. F., BAIRD K. M. Mutation to extended host range and the occurrence of phenotypic mixing in the temperate coliphage lambda. Virology. 1956 Aug;2(4):565–574. doi: 10.1016/0042-6822(56)90012-5. [DOI] [PubMed] [Google Scholar]
  2. Adams J. M., Cory S., Spahr P. F. Nucleotide sequences of fragments of R17 bacteriophage RNA from the region immediately preceding the coat-protein cistron. Eur J Biochem. 1972 Sep 25;29(3):469–479. doi: 10.1111/j.1432-1033.1972.tb02011.x. [DOI] [PubMed] [Google Scholar]
  3. Atkins J. F., Gesteland R. F., Reid B. R., Anderson C. W. Normal tRNAs promote ribosomal frameshifting. Cell. 1979 Dec;18(4):1119–1131. doi: 10.1016/0092-8674(79)90225-3. [DOI] [PubMed] [Google Scholar]
  4. Bolivar F., Rodriguez R. L., Greene P. J., Betlach M. C., Heyneker H. L., Boyer H. W., Crosa J. H., Falkow S. Construction and characterization of new cloning vehicles. II. A multipurpose cloning system. Gene. 1977;2(2):95–113. [PubMed] [Google Scholar]
  5. Devos R., Contreras R., van Emmelo J., Fiers W. Identification of the translocatable element IS1 in a molecular chimera constructed with plasmid pBR322 DNA into which a bacteriophage MS2 DNA copy was inserted by the poly(dA).poly(dT) linker method. J Mol Biol. 1979 Mar 15;128(4):621–632. doi: 10.1016/0022-2836(79)90296-1. [DOI] [PubMed] [Google Scholar]
  6. Devos R., van Emmelo J., Contreras R., Fiers W. Construction and characterization of a plasmid containing a nearly full-size DNA copy of bacteriophage MS2 RNA. J Mol Biol. 1979 Mar 15;128(4):595–619. doi: 10.1016/0022-2836(79)90295-x. [DOI] [PubMed] [Google Scholar]
  7. Dürwald H., Hoffmann-Berling H. Endonuclease-I-deficient and ribonuclease I-deficient Escherichia coli mutants. J Mol Biol. 1968 Jul 14;34(2):331–346. doi: 10.1016/0022-2836(68)90257-x. [DOI] [PubMed] [Google Scholar]
  8. Fiers W., Contreras R., Duerinck F., Haegeman G., Iserentant D., Merregaert J., Min Jou W., Molemans F., Raeymaekers A., Van den Berghe A. Complete nucleotide sequence of bacteriophage MS2 RNA: primary and secondary structure of the replicase gene. Nature. 1976 Apr 8;260(5551):500–507. doi: 10.1038/260500a0. [DOI] [PubMed] [Google Scholar]
  9. Fiers W., Contreras R., Duerinck F., Haegmean G., Merregaert J., Jou W. M., Raeymakers A., Volckaert G., Ysebaert M., Van de Kerckhove J. A-protein gene of bacteriophage MS2. Nature. 1975 Jul 24;256(5515):273–278. doi: 10.1038/256273a0. [DOI] [PubMed] [Google Scholar]
  10. Humayun M. Z., Chambers R. W. Construction and characterization of an Escherichia coli plasmid bearing a functional gene G of bacteriophage phiX174. Proc Natl Acad Sci U S A. 1978 Feb;75(2):774–778. doi: 10.1073/pnas.75.2.774. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Kahn M., Kolter R., Thomas C., Figurski D., Meyer R., Remaut E., Helinski D. R. Plasmid cloning vehicles derived from plasmids ColE1, F, R6K, and RK2. Methods Enzymol. 1979;68:268–280. doi: 10.1016/0076-6879(79)68019-9. [DOI] [PubMed] [Google Scholar]
  12. Kamen R. Characterization of the subunits of Q-beta replicase. Nature. 1970 Nov 7;228(5271):527–533. doi: 10.1038/228527a0. [DOI] [PubMed] [Google Scholar]
  13. Kastelein R. A., Remaut E., Fiers W., van Duin J. Lysis gene expression of RNA phage MS2 depends on a frameshift during translation of the overlapping coat protein gene. Nature. 1982 Jan 7;295(5844):35–41. doi: 10.1038/295035a0. [DOI] [PubMed] [Google Scholar]
  14. Kondo M., Gallerani R., Weissmann C. Subunit structure of Q-beta replicase. Nature. 1970 Nov 7;228(5271):525–527. doi: 10.1038/228525a0. [DOI] [PubMed] [Google Scholar]
  15. Kozak M., Nathans D. Translation of the genome of a ribonucleic acid bacteriophage. Bacteriol Rev. 1972 Mar;36(1):109–134. doi: 10.1128/br.36.1.109-134.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. LOEB T., ZINDER N. D. A bacteriophage containing RNA. Proc Natl Acad Sci U S A. 1961 Mar 15;47:282–289. doi: 10.1073/pnas.47.3.282. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  18. Loomis W. F., Jr, Magasanik B. The catabolite repression gene of the lac operon in Escherichia coli. J Mol Biol. 1967 Feb 14;23(3):487–494. doi: 10.1016/s0022-2836(67)80120-7. [DOI] [PubMed] [Google Scholar]
  19. Porter A. G., Hindley J. The binding of Q initiator fragments to E. coli ribosomes. FEBS Lett. 1973 Jul 15;33(3):339–342. doi: 10.1016/0014-5793(73)80225-x. [DOI] [PubMed] [Google Scholar]
  20. Remaut E., Fiers W. Studies on the bacteriophage MS2. XVI. The termination signal of the A protein cistron. J Mol Biol. 1972 Nov 14;71(2):243–261. doi: 10.1016/0022-2836(72)90349-x. [DOI] [PubMed] [Google Scholar]
  21. Remaut E., Stanssens P., Fiers W. Plasmid vectors for high-efficiency expression controlled by the PL promoter of coliphage lambda. Gene. 1981 Oct;15(1):81–93. doi: 10.1016/0378-1119(81)90106-2. [DOI] [PubMed] [Google Scholar]
  22. Robertson H. D., Lodish H. F. Messenger characteristics of nascent bacteriophage RNA. Proc Natl Acad Sci U S A. 1970 Oct;67(2):710–716. doi: 10.1073/pnas.67.2.710. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Sekizawa J., Inouye S., Halegoua S., Inouye M. Precursors of major outer membrane proteins of Escherichia coli. Biochem Biophys Res Commun. 1977 Aug 8;77(3):1126–1133. doi: 10.1016/s0006-291x(77)80095-8. [DOI] [PubMed] [Google Scholar]
  24. Skalka A., Shapiro L. In situ immunoassays for gene translation products in phage plaques and bacterial colonies. Gene. 1976;1(1):65–79. doi: 10.1016/0378-1119(76)90007-x. [DOI] [PubMed] [Google Scholar]
  25. Steitz J. A. Discriminatory ribosome rebinding of isolated regions of protein synthesis initiation from the ribonucleic acid of bacteriophage R17. Proc Natl Acad Sci U S A. 1973 Sep;70(9):2605–2609. doi: 10.1073/pnas.70.9.2605. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Sugiyama T., Stone H. O., Jr Viral and bacterial protein syntheses in MS2-infected Escherichia coli cells. J Mol Biol. 1968 Aug 28;36(1):91–105. doi: 10.1016/0022-2836(68)90222-2. [DOI] [PubMed] [Google Scholar]
  27. 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]
  28. Tooze J., Weber K. Isolation and characterization of amber mutants of bacteriophage R17. J Mol Biol. 1967 Sep 14;28(2):311–330. doi: 10.1016/s0022-2836(67)80012-3. [DOI] [PubMed] [Google Scholar]
  29. Vandamme E., Remaut E., van Montagu M., Fiers W. Studies on the bacteriophage MS 2. XVII. Suppressor-sensitive mutants of the A protein cistron. Mol Gen Genet. 1972;117(3):219–228. doi: 10.1007/BF00271649. [DOI] [PubMed] [Google Scholar]
  30. Wahba A. J., Miller M. J., Niveleau A., Landers T. A., Carmichael G. G., Weber K., Hawley D. A., Slobin L. I. Subunit I of G beta replicase and 30 S ribosomal protein S1 of Escherichia coli. Evidence for the identity of the two proteins. J Biol Chem. 1974 May 25;249(10):3314–3316. [PubMed] [Google Scholar]

Articles from The EMBO Journal are provided here courtesy of Nature Publishing Group

RESOURCES