Abstract
Translocation of bacteriophage T7 DNA from the capsid into the cell has been assayed by measuring the time after infection that each GATC site on the phage genome is methylated by cells containing high levels of DNA adenine methylase. Methylation at GATC sites on T7 DNA renders both the infecting genome and any newly synthesized molecules sensitive to the restriction enzyme DpnI. In a normal infection at 30 degrees C, translocation of the T7 genome into the cell takes between 9 and 12 min. In contrast, translocation of the entire phage lambda genome or of a T7 genome ejected from a lambda capsid can be detected within the first minute of infection. Entry of the leading end of the T7 genome occurs by a transcription-independent mechanism that brings both Escherichia coli and T7 promoters into the cell. Further translocation of the genome normally involves transcription by the RNA polymerases of both E. coli and T7; the rates of DNA translocation into the cell when catalyzed by each enzyme are comparable to the estimated rates of transcription of the respective enzymes. A GATC site located between the early E. coli promoters and the coding sequences of the first T7 protein made after infection is not methylated before the protein is synthesized, a result supporting the idea (B. A. Moffatt and F. W. Studier, J. Bacteriol. 170:2095-2105, 1988) that only certain proteins are permitted access to the entering T7 DNA. In the absence of transcription, the genomes of most T7 strains do not completely enter the cell. However, the entire genome of a mutant that lacks bp 3936 to 808 of T7 DNA enters the cell in a transcription-independent process at an average overall rate of 50 bp per s.
Full Text
The Full Text of this article is available as a PDF (871.3 KB).
Selected References
These references are in PubMed. This may not be the complete list of references from this article.
- Beck P. J., Molineux I. J. Defective transcription of the right end of bacteriophage T7 DNA during an abortive infection of F plasmid-containing Escherichia coli. J Bacteriol. 1991 Feb;173(3):947–954. doi: 10.1128/jb.173.3.947-954.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Bremer H., Yuan D. RNA chain growth-rate in Escherichia coli. J Mol Biol. 1968 Dec 14;38(2):163–180. doi: 10.1016/0022-2836(68)90404-x. [DOI] [PubMed] [Google Scholar]
- 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]
- Brunovskis I., Summers W. C. The process of infection with coliphage T7. V. Shutoff of host RNA synthesis by an early phage function. Virology. 1971 Jul;45(1):224–231. doi: 10.1016/0042-6822(71)90129-2. [DOI] [PubMed] [Google Scholar]
- Casjens S., Adams M. B. Posttranscriptional modulation of bacteriophage P22 scaffolding protein gene expression. J Virol. 1985 Jan;53(1):185–191. doi: 10.1128/jvi.53.1.185-191.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Chapman K. A., Burgess R. R. Construction of bacteriophage T7 late promoters with point mutations and characterization by in vitro transcription properties. Nucleic Acids Res. 1987 Jul 10;15(13):5413–5432. doi: 10.1093/nar/15.13.5413. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Chattoraj D. K., Inman R. B. Location of DNA ends in P2, 186, P4 and lambda bacteriophage heads. J Mol Biol. 1974 Jul 25;87(1):11–22. doi: 10.1016/0022-2836(74)90556-7. [DOI] [PubMed] [Google Scholar]
- Condreay J. P., Molineux I. J. Synthesis of the capsid protein inhibits development of bacteriophage T3 mutants that abortively infect F plasmid-containing cells. J Mol Biol. 1989 Jun 5;207(3):543–554. doi: 10.1016/0022-2836(89)90463-4. [DOI] [PubMed] [Google Scholar]
- Condreay J. P., Wright S. E., Molineux I. J. Nucleotide sequence and complementation studies of the gene 10 region of bacteriophage T3. J Mol Biol. 1989 Jun 5;207(3):555–561. doi: 10.1016/0022-2836(89)90464-6. [DOI] [PubMed] [Google Scholar]
- Davanloo P., Rosenberg A. H., Dunn J. J., Studier F. W. Cloning and expression of the gene for bacteriophage T7 RNA polymerase. Proc Natl Acad Sci U S A. 1984 Apr;81(7):2035–2039. doi: 10.1073/pnas.81.7.2035. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Dunn J. J., Studier F. W. Complete nucleotide sequence of bacteriophage T7 DNA and the locations of T7 genetic elements. J Mol Biol. 1983 Jun 5;166(4):477–535. doi: 10.1016/s0022-2836(83)80282-4. [DOI] [PubMed] [Google Scholar]
- García L. R., Molineux I. J. Incomplete entry of bacteriophage T7 DNA into F plasmid-containing Escherichia coli. J Bacteriol. 1995 Jul;177(14):4077–4083. doi: 10.1128/jb.177.14.4077-4083.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Ikeda R. A., Lin A. C., Clarke J. Initiation of transcription by T7 RNA polymerase as its natural promoters. J Biol Chem. 1992 Feb 5;267(4):2640–2649. [PubMed] [Google Scholar]
- Manor H., Goodman D., Stent G. S. RNA chain growth rates in Escherichia coli. J Mol Biol. 1969 Jan 14;39(1):1–29. doi: 10.1016/0022-2836(69)90329-5. [DOI] [PubMed] [Google Scholar]
- Marinus M. G., Poteete A., Arraj J. A. Correlation of DNA adenine methylase activity with spontaneous mutability in Escherichia coli K-12. Gene. 1984 Apr;28(1):123–125. doi: 10.1016/0378-1119(84)90095-7. [DOI] [PubMed] [Google Scholar]
- McAllister W. T., Barrett C. L. Hybridization mapping of restriction fragments from the early region of bacteriophage T7 DNA. Virology. 1977 Oct 15;82(2):275–287. doi: 10.1016/0042-6822(77)90003-4. [DOI] [PubMed] [Google Scholar]
- McAllister W. T., McCarron R. J. Hybridization of the in vitro products of bacteriop&hage T7 RNA polymerase to restriction fragments of T7 DNA. Virology. 1977 Oct 15;82(2):288–298. doi: 10.1016/0042-6822(77)90004-6. [DOI] [PubMed] [Google Scholar]
- McAllister W. T., Morris C., Rosenberg A. H., Studier F. W. Utilization of bacteriophage T7 late promoters in recombinant plasmids during infection. J Mol Biol. 1981 Dec 15;153(3):527–544. doi: 10.1016/0022-2836(81)90406-x. [DOI] [PubMed] [Google Scholar]
- McAllister W. T., Wu H. L. Regulation of transcription of the late genes of bacteriophage T7. Proc Natl Acad Sci U S A. 1978 Feb;75(2):804–808. doi: 10.1073/pnas.75.2.804. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Moffatt B. A., Studier F. W. Entry of bacteriophage T7 DNA into the cell and escape from host restriction. J Bacteriol. 1988 May;170(5):2095–2105. doi: 10.1128/jb.170.5.2095-2105.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Moffatt B. A., Studier F. W. T7 lysozyme inhibits transcription by T7 RNA polymerase. Cell. 1987 Apr 24;49(2):221–227. doi: 10.1016/0092-8674(87)90563-0. [DOI] [PubMed] [Google Scholar]
- Morrison T. G., Blumberg D. D., Malamy M. H. T7 protein synthesis in F' episome-containing cells: assignment of specific proteins to three translational groups. J Virol. 1974 Feb;13(2):386–393. doi: 10.1128/jvi.13.2.386-393.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Pao C. C., Speyer J. F. Order of injection of T7 bacteriophage DNA. J Virol. 1973 Jun;11(6):1024–1026. doi: 10.1128/jvi.11.6.1024-1026.1973. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Roessner C. A., Ihler G. M. Proteinase sensitivity of bacteriophage lambda tail proteins gpJ and pH in complexes with the lambda receptor. J Bacteriol. 1984 Jan;157(1):165–170. doi: 10.1128/jb.157.1.165-170.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Rose J. K., Mosteller R. D., Yanofsky C. Tryptophan messenger ribonucleic acid elongation rates and steady-state levels of tryptophan operon enzymes under various growth conditions. J Mol Biol. 1970 Aug;51(3):541–550. doi: 10.1016/0022-2836(70)90007-0. [DOI] [PubMed] [Google Scholar]
- Rosenberg S. M., Stahl M. M., Kobayashi I., Stahl F. W. Improved in vitro packaging of coliphage lambda DNA: a one-strain system free from endogenous phage. Gene. 1985;38(1-3):165–175. doi: 10.1016/0378-1119(85)90215-x. [DOI] [PubMed] [Google Scholar]
- Saigo K. Polar DNA ejection in bacteriophage T7. Virology. 1975 May;65(1):120–127. doi: 10.1016/0042-6822(75)90012-4. [DOI] [PubMed] [Google Scholar]
- Saigo K., Uchida H. Connection of the right-hand terminus of DNA to the proximal end of the tail in bacteriophage lambda. Virology. 1974 Oct;61(2):524–536. doi: 10.1016/0042-6822(74)90287-6. [DOI] [PubMed] [Google Scholar]
- Schmitt C. K., Kemp P., Molineux I. J. Genes 1.2 and 10 of bacteriophages T3 and T7 determine the permeability lesions observed in infected cells of Escherichia coli expressing the F plasmid gene pifA. J Bacteriol. 1991 Oct;173(20):6507–6514. doi: 10.1128/jb.173.20.6507-6514.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Shinder G., Gold M. The Nul subunit of bacteriophage lambda terminase binds to specific sites in cos DNA. J Virol. 1988 Feb;62(2):387–392. doi: 10.1128/jvi.62.2.387-392.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Strome S., Young E. T. Chemical and functional quantitation of gene 0.3 messenger RNA during T7 infection. J Mol Biol. 1980 Feb 5;136(4):417–432. doi: 10.1016/0022-2836(80)90398-8. [DOI] [PubMed] [Google Scholar]
- Strome S., Young E. T. Translational control of the expression of bacteriophage T7 gene 0.3. J Mol Biol. 1978 Oct 15;125(1):75–93. doi: 10.1016/0022-2836(78)90255-3. [DOI] [PubMed] [Google Scholar]
- Strome S., Young E. T. Translational discrimination against bacteriophage T7 gene 0.3 messenger RNA. J Mol Biol. 1980 Feb 5;136(4):433–450. doi: 10.1016/0022-2836(80)90399-x. [DOI] [PubMed] [Google Scholar]
- Studier F. W. Bacteriophage T7. Science. 1972 Apr 28;176(4033):367–376. doi: 10.1126/science.176.4033.367. [DOI] [PubMed] [Google Scholar]
- Studier F. W. Genetic mapping of a mutation that causes ribonucleases III deficiency in Escherichia coli. J Bacteriol. 1975 Oct;124(1):307–316. doi: 10.1128/jb.124.1.307-316.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Studier F. W., Rosenberg A. H. Genetic and physical mapping of the late region of bacteriophage T7 DNA by use of cloned fragments of T7 DNA. J Mol Biol. 1981 Dec 15;153(3):503–525. doi: 10.1016/0022-2836(81)90405-8. [DOI] [PubMed] [Google Scholar]
- Thomas J. O. Chemical linkage of the tail to the right-hand end of bacteriophage lambda DNA. J Mol Biol. 1974 Jul 25;87(1):1–9. doi: 10.1016/0022-2836(74)90555-5. [DOI] [PubMed] [Google Scholar]
- Xu S. Y., Feiss M. The last duplex base-pair of the phage lambda chromosome. Involvement in packaging, ejection and routing of lambda DNA. J Mol Biol. 1991 Jul 20;220(2):293–306. doi: 10.1016/0022-2836(91)90014-w. [DOI] [PubMed] [Google Scholar]
- Zavriev S. K., Shemyakin M. F. RNA polymerase-dependent mechanism for the stepwise T7 phage DNA transport from the virion into E. coli. Nucleic Acids Res. 1982 Mar 11;10(5):1635–1652. doi: 10.1093/nar/10.5.1635. [DOI] [PMC free article] [PubMed] [Google Scholar]