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. 1976 Feb;17(2):642–658. doi: 10.1128/jvi.17.2.642-658.1976

Early gene expression in bacteriophage T7. I. In vivo synthesis, inactivation, and translational utilization of early mRNA's.

K Hercules, S Jovanovich, W Sauerbrier
PMCID: PMC515454  PMID: 1255850

Abstract

In vivo decay rates for the individual T7 early mRNA species were determined. The physical half-lives, measured at 37 C, range from 1.1 min for gene 0.7 RNA to 4.5 min for gene 0.3 RNA. Physical half-lives, as observed after rifampin inhibition of RNA synthesis and polyacylamide electrophoresis of RNAs, are approximately 30% longer than functional half-lives, as observed by 14C-labeled amino acid uptake into individual T7 early proteins. The different RNA species are synthesized at grossly different rates, 0.3 RNA at four times the rate of 1.0 RNA, 0.7 RNA at twice the rate, and 1.1 and 1.3 RNAs at about the same or a slightly lower rate than 1.0 RNA. Rho-factor-mediated termination of transcription behind genes 0.3, 0.7, and perhaps behind 1.0 is inferred from these data. The in vivo translational utilization of the individual T7 early-message species was found to vary by not more than a factor of 2.

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Selected References

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  1. Achord D., Kennell D. Metabolism of messenger RNA from the gal operon of Escherichia coli. J Mol Biol. 1974 Dec 15;90(3):581–599. doi: 10.1016/0022-2836(74)90236-8. [DOI] [PubMed] [Google Scholar]
  2. Apirion D. Degradation of RNA in Escherichia coli. A hypothesis. Mol Gen Genet. 1973 May 28;122(4):313–322. doi: 10.1007/BF00269431. [DOI] [PubMed] [Google Scholar]
  3. Apirion D., Watson N. Analysis of an Escherichia coli strain carrying physiologically compensating mutations one of which causes an altered ribonuclease 3. Mol Gen Genet. 1974;132(2):89–104. doi: 10.1007/BF00272175. [DOI] [PubMed] [Google Scholar]
  4. Arrand J. R., Hindley J. Nucleotide sequence of a ribosome binding site on RNA synthesized in vitro from coliphage T7. Nat New Biol. 1973 Jul 4;244(131):10–13. doi: 10.1038/newbio244010a0. [DOI] [PubMed] [Google Scholar]
  5. Blundell M., Kennell D. Evidence for endonucleolytic attack in decay of lac messenger RNA in Escherichia coli. J Mol Biol. 1974 Feb 25;83(2):143–161. doi: 10.1016/0022-2836(74)90385-4. [DOI] [PubMed] [Google Scholar]
  6. Both G. W., Banerjee A. K., Shatkin A. J. Methylation-dependent translation of viral messenger RNAs in vitro. Proc Natl Acad Sci U S A. 1975 Mar;72(3):1189–1193. doi: 10.1073/pnas.72.3.1189. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Brunovskis I., Summers W. C. The process of infection with coliphage 17. VI. A phage gene controlling shutoff of host RNA synthesis. Virology. 1972 Nov;50(2):322–327. doi: 10.1016/0042-6822(72)90383-2. [DOI] [PubMed] [Google Scholar]
  8. Bräutigam A. R., Sauerbier W. Transcription unit mapping in bacteriophage T7. I. In vivo transcription by Escherichia coli RNA polymerase. J Virol. 1973 Oct;12(4):882–886. doi: 10.1128/jvi.12.4.882-886.1973. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Darlix J. L., Horaist M. Existence and possible roles of transcriptional barriers in T7 DNA early region as shown by electron microscopy. Nature. 1975 Jul 24;256(5515):288–292. doi: 10.1038/256288a0. [DOI] [PubMed] [Google Scholar]
  10. Darlix J. L. Rho, a factor causing the modulation of early T7 genes transcription. Biochimie. 1974;56(5):693–701. doi: 10.1016/s0300-9084(74)80040-4. [DOI] [PubMed] [Google Scholar]
  11. Dunn J. J., Studier F. W. T7 early RNAs and Escherichia coli ribosomal RNAs are cut from large precursor RNAs in vivo by ribonuclease 3. Proc Natl Acad Sci U S A. 1973 Dec;70(12):3296–3300. doi: 10.1073/pnas.70.12.3296. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Dunn J. J., Studier F. W. T7 early RNAs are generated by site-specific cleavages. Proc Natl Acad Sci U S A. 1973 May;70(5):1559–1563. doi: 10.1073/pnas.70.5.1559. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Gupta R. S., Schlessinger D. Differential modes of chemical decay for early and late lambda messenger RNA. J Mol Biol. 1975 Feb 25;92(2):311–318. doi: 10.1016/0022-2836(75)90230-2. [DOI] [PubMed] [Google Scholar]
  14. Gupta S. L., Chen J., Schaefer L., Lengyel P., Weissman S. M. Nucleotide sequence of a ribosome attachment site of bacteriophage f2 RNA. Biochem Biophys Res Commun. 1970 Jun 5;39(5):883–888. doi: 10.1016/0006-291x(70)90406-7. [DOI] [PubMed] [Google Scholar]
  15. HILL R. F. A radiation-sensitive mutant of Escherichia coli. Biochim Biophys Acta. 1958 Dec;30(3):636–637. doi: 10.1016/0006-3002(58)90112-4. [DOI] [PubMed] [Google Scholar]
  16. Hercules K., Schweiger M., Sauerbier W. Cleavage by RNase 3 converts T3 and T7 early precursor RNA into translatable message. Proc Natl Acad Sci U S A. 1974 Mar;71(3):840–844. doi: 10.1073/pnas.71.3.840. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Herrlich P., Rahmsdorf H. J., Pai S. H., Schweigher M. Translational control induced by bacteriophage T7. Proc Natl Acad Sci U S A. 1974 Apr;71(4):1088–1092. doi: 10.1073/pnas.71.4.1088. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Hopper J. E., Ko G., Young E. T. Comparative analysis of the in vivo and in vitro expression of bacteriophage T7 messenger RNAs during infection of Escherichia coli. J Mol Biol. 1975 Jun 5;94(4):539–554. doi: 10.1016/0022-2836(75)90320-4. [DOI] [PubMed] [Google Scholar]
  19. Karam J. D., Bowles M. G. Mutation to overproduction of bacteriophage T4 gene products. J Virol. 1974 Feb;13(2):428–438. doi: 10.1128/jvi.13.2.428-438.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Kennell D., Bicknell I. Decay of messenger ribonucleic acid from the lactose operon of Escherichia coli as a function of growth temperature. J Mol Biol. 1973 Feb 15;74(1):21–31. doi: 10.1016/0022-2836(73)90351-3. [DOI] [PubMed] [Google Scholar]
  21. Kindler P., Keil T. U., Hofschneider P. H. Isolation and characterization of a ribonuclease 3 deficient mutant of Escherichia coli. Mol Gen Genet. 1973 Oct 16;126(1):53–59. doi: 10.1007/BF00333481. [DOI] [PubMed] [Google Scholar]
  22. Loening U. E. The fractionation of high-molecular-weight ribonucleic acid by polyacrylamide-gel electrophoresis. Biochem J. 1967 Jan;102(1):251–257. doi: 10.1042/bj1020251. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Maizels N. E. coli lactose operon ribosome binding site. Nature. 1974 Jun 14;249(458):647–649. doi: 10.1038/249647b0. [DOI] [PubMed] [Google Scholar]
  24. Marrs B. L., Yanofsky C. Host and bacteriophage specific messenger RNA degradation in T7-infected Escherichia coli. Nat New Biol. 1971 Dec 8;234(49):168–170. doi: 10.1038/newbio234168a0. [DOI] [PubMed] [Google Scholar]
  25. Minkley E. G., Jr Transcription of the early region of bacteriophage T7: characterization of the in vivo transcripts. J Mol Biol. 1974 Mar;83(3):289–304. doi: 10.1016/0022-2836(74)90281-2. [DOI] [PubMed] [Google Scholar]
  26. Nikolaev N., Silengo L., Schlessinger D. A role for ribonuclease 3 in processing of ribosomal ribonucleic acid and messenger ribonucleic acid precursors in Escherichia coli. J Biol Chem. 1973 Nov 25;248(22):7967–7969. [PubMed] [Google Scholar]
  27. Ponta H., Rahmsdorf H. J., Pai S. H., Hirsch-Kauffmann M., Herrlich P., Schweiger M. Control of gene expression in bacteriophage T7: transcriptional controls. Mol Gen Genet. 1974;134(4):281–297. doi: 10.1007/BF00337463. [DOI] [PubMed] [Google Scholar]
  28. Puga A., Borrás M. T., Tessman E. S., Tessman I. Difference between functional and structural integrity of messenger RNA. Proc Natl Acad Sci U S A. 1973 Jul;70(7):2171–2175. doi: 10.1073/pnas.70.7.2171. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Rahmsdorf H. J., Pai S. H., Ponta H., Herrlich P., Roskoski R., Jr, Schweiger M., Studier F. W. Protein kinase induction in Escherichia coli by bacteriophage T7. Proc Natl Acad Sci U S A. 1974 Feb;71(2):586–589. doi: 10.1073/pnas.71.2.586. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Robertson H. D., Webster R. E., Zinder N. D. Purification and properties of ribonuclease III from Escherichia coli. J Biol Chem. 1968 Jan 10;243(1):82–91. [PubMed] [Google Scholar]
  31. Rosenberg M., Kramer R. A., Steitz J. A. T7 early messenger RNAs are the direct products of ribonuclease III cleavage. J Mol Biol. 1974 Nov 15;89(4):777–782. doi: 10.1016/0022-2836(74)90052-7. [DOI] [PubMed] [Google Scholar]
  32. Sauerbier W., Hercules K. Control of gene function in bacteriophage T4. IV. Post-transcriptional shutoff of expression of early genes. J Virol. 1973 Sep;12(3):538–547. doi: 10.1128/jvi.12.3.538-547.1973. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Schleicher M., Bautz E. K.F. Stability of phage T7 gene I mRNA in E. coli cells infected with wild type phage and with gene I amber mutants. FEBS Lett. 1972 Dec 1;28(2):139–141. doi: 10.1016/0014-5793(72)80695-1. [DOI] [PubMed] [Google Scholar]
  34. Schweiger M., Herrlich P., Scherzinger E., Rahmsdorf H. J. Negative control of protein synthesis after infection with bacteriophage T7. Proc Natl Acad Sci U S A. 1972 Aug;69(8):2203–2207. doi: 10.1073/pnas.69.8.2203. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Shine J., Dalgarno L. Determinant of cistron specificity in bacterial ribosomes. Nature. 1975 Mar 6;254(5495):34–38. doi: 10.1038/254034a0. [DOI] [PubMed] [Google Scholar]
  36. Siegel R. B., Summers W. C. The process of infection with coliphage T7. 3. Control of phage-specific RNA synthesis in vivo by an early phage gene. J Mol Biol. 1970 Apr 14;49(1):115–123. doi: 10.1016/0022-2836(70)90380-3. [DOI] [PubMed] [Google Scholar]
  37. Simon M. N., Studier F. W. Physical mapping of the early region of bacteriophage T7 DNA. J Mol Biol. 1973 Sep 15;79(2):249–265. doi: 10.1016/0022-2836(73)90004-1. [DOI] [PubMed] [Google Scholar]
  38. Staples D. H., Hindley J., Billeter M. A., Weissmann C. Localization of Q-beta maturation cistron ribosome binding site. Nat New Biol. 1971 Sep 15;234(50):202–204. doi: 10.1038/newbio234202a0. [DOI] [PubMed] [Google Scholar]
  39. 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]
  40. Studier F. W. Analysis of bacteriophage T7 early RNAs and proteins on slab gels. J Mol Biol. 1973 Sep 15;79(2):237–248. doi: 10.1016/0022-2836(73)90003-x. [DOI] [PubMed] [Google Scholar]
  41. Studier F. W. Genetic analysis of non-essential bacteriophage T7 genes. J Mol Biol. 1973 Sep 15;79(2):227–236. doi: 10.1016/0022-2836(73)90002-8. [DOI] [PubMed] [Google Scholar]
  42. Summers W. C., Brunovskis I., Hyman R. W. The process of infection with coliphage T7. VII. Characterization and mapping of the major in vivo transcription products of the early region. J Mol Biol. 1973 Mar 5;74(3):291–300. doi: 10.1016/0022-2836(73)90374-4. [DOI] [PubMed] [Google Scholar]
  43. Summers W. C. The process of infection with coliphage T7. I. Characterization of T7 RNA by polyacrylamide gel electrophoretic analysis. Virology. 1969 Oct;39(2):175–181. doi: 10.1016/0042-6822(69)90037-3. [DOI] [PubMed] [Google Scholar]
  44. Summers W. C. The process of infection with coliphage T7. IV. Stability of RNA in bacteriophage-infected cells. J Mol Biol. 1970 Aug;51(3):671–678. doi: 10.1016/0022-2836(70)90015-x. [DOI] [PubMed] [Google Scholar]
  45. Vollenweider H. J., Sogo J. M., Koller T. A routine method for protein-free spreading of double- and single-stranded nucleic acid molecules. Proc Natl Acad Sci U S A. 1975 Jan;72(1):83–87. doi: 10.1073/pnas.72.1.83. [DOI] [PMC free article] [PubMed] [Google Scholar]
  46. Wiberg J. S., Mendelsohn S., Warner V., Hercules K., Aldrich C., Munro J. L. SP62, a viable mutant of bacteriophage T4D defective in regulation of phage enzyme synthesis. J Virol. 1973 Oct;12(4):775–792. doi: 10.1128/jvi.12.4.775-792.1973. [DOI] [PMC free article] [PubMed] [Google Scholar]
  47. Yamamoto T., Imamoto F. Differential stability of trp messenger RNA synthesized originating at the trp promoter and pL promoter of lambda trp phage. J Mol Biol. 1975 Feb 25;92(2):289–304. doi: 10.1016/0022-2836(75)90228-4. [DOI] [PubMed] [Google Scholar]
  48. Zillig W., Fujiki H., Blum W., Janeković D., Schweiger M., Rahmsdorf H., Ponta H., Hirsch-Kauffmann M. In vivo and in vitro phosphorylation of DNA-dependent RNA polymerase of Escherichia coli by bacteriophage-T7-induced protein kinase. Proc Natl Acad Sci U S A. 1975 Jul;72(7):2506–2510. doi: 10.1073/pnas.72.7.2506. [DOI] [PMC free article] [PubMed] [Google Scholar]

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