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
. 1984 Dec;81(23):7402–7406. doi: 10.1073/pnas.81.23.7402

Multiple initiation sites of DNA replication flanking the origin region of lambda dv genome.

T Tsurimoto, K Matsubara
PMCID: PMC392154  PMID: 6095292

Abstract

Early replicative intermediates of lambda dv plasmid were prepared by an in vitro replication system in the presence of 2',3'-dideoxycytidine 5'-triphosphate, an inhibitor of DNA chain elongation. Short-chain DNAs produced from regions near the replication origin were purified from the intermediates. A fraction of the DNAs was covalently linked to primer RNA. The transition sites from primer RNA to DNA synthesis were mapped along the nucleotide sequence of the genome, by eliminating the RNA by alkaline hydrolysis and labeling the freshly exposed 5' ends of DNA with 32P. The transition sites were found to be located on both sides of the ori region, which includes four 19-base-pair repeats where one of the lambda specific initiator proteins, O, binds. No transition arose within the ori region. The transition sites are multiple on both sides of the ori region and are clustered in one of the two strands in such a way that DNA syntheses from the two sides converge. The frequency of the "leftward" DNA synthesis is several times higher than that of "rightward" synthesis, reflecting the asymmetric bidirectional replication of lambda dv DNA.

Full text

PDF
7406

Images in this article

7404Image
on p.7404
7404Image
on p.7404

Selected References

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

  1. Chow L. T., Davidson N. Electron microscope study of the structures of lambdadv DNAs. J Mol Biol. 1974 Jun 15;86(1):69–89. doi: 10.1016/s0022-2836(74)80008-2. [DOI] [PubMed] [Google Scholar]
  2. Denniston-Thompson K., Moore D. D., Kruger K. E., Furth M. E., Blattner F. R. Physical structure of the replication origin of bacteriophage lambda. Science. 1977 Dec 9;198(4321):1051–1056. doi: 10.1126/science.929187. [DOI] [PubMed] [Google Scholar]
  3. Fiddes J. C., Barrell B. G., Godson G. N. Nucleotide sequences of the separate origins of synthesis of bacteriophage G4 viral and complementary DNA strands. Proc Natl Acad Sci U S A. 1978 Mar;75(3):1081–1085. doi: 10.1073/pnas.75.3.1081. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Fujiyama A., Kohara Y., Okazaki T. Initiation sites for discontinuous DNA synthesis of bacteriophage T7. Proc Natl Acad Sci U S A. 1981 Feb;78(2):903–907. doi: 10.1073/pnas.78.2.903. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Fuller R. S., Kaguni J. M., Kornberg A. Enzymatic replication of the origin of the Escherichia coli chromosome. Proc Natl Acad Sci U S A. 1981 Dec;78(12):7370–7374. doi: 10.1073/pnas.78.12.7370. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Grosschedl R., Hobom G. DNA sequences and structural homologies of the replication origins of lambdoid bacteriophages. Nature. 1979 Feb 22;277(5698):621–627. doi: 10.1038/277621a0. [DOI] [PubMed] [Google Scholar]
  7. Hay R. T., DePamphilis M. L. Initiation of SV40 DNA replication in vivo: location and structure of 5' ends of DNA synthesized in the ori region. Cell. 1982 Apr;28(4):767–779. doi: 10.1016/0092-8674(82)90056-3. [DOI] [PubMed] [Google Scholar]
  8. Itoh T., Tomizawa J. Formation of an RNA primer for initiation of replication of ColE1 DNA by ribonuclease H. Proc Natl Acad Sci U S A. 1980 May;77(5):2450–2454. doi: 10.1073/pnas.77.5.2450. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Kiger J. A., Jr, Sinsheimer R. L. Vegetative lambda DNA. IV. Fractionation of replicating lambda DNA on benzoylated-naphthoylated DEAE cellulose. J Mol Biol. 1969 Mar 28;40(3):467–490. doi: 10.1016/0022-2836(69)90166-1. [DOI] [PubMed] [Google Scholar]
  10. Matsubara K. Replication control system in lambda dv. Plasmid. 1981 Jan;5(1):32–52. doi: 10.1016/0147-619x(81)90076-7. [DOI] [PubMed] [Google Scholar]
  11. Messing J., Vieira J. A new pair of M13 vectors for selecting either DNA strand of double-digest restriction fragments. Gene. 1982 Oct;19(3):269–276. doi: 10.1016/0378-1119(82)90016-6. [DOI] [PubMed] [Google Scholar]
  12. Ogawa T., Okazaki T. RNA-linked nascent DNA pieces in phage T7-infected Escherichia coli. III. Detection of intact primer RNA. Nucleic Acids Res. 1979 Nov 24;7(6):1621–1633. doi: 10.1093/nar/7.6.1621. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Ogawa T., Tomizawa J. Replication of bacteriophage DNA. I. Replication of DNA of lambda phage defective in early functions. J Mol Biol. 1968 Dec 14;38(2):217–225. doi: 10.1016/0022-2836(68)90407-5. [DOI] [PubMed] [Google Scholar]
  14. Okazaki R., Hirose S., Okazaki T., Ogawa T., Kurosawa Y. Assay of RNA-linked nascent DNA pieces with polynucleotide kinase. Biochem Biophys Res Commun. 1975 Feb 17;62(4):1018–1024. doi: 10.1016/0006-291x(75)90424-6. [DOI] [PubMed] [Google Scholar]
  15. Rambach A. Replicator mutants of bacteriophage lambda: characterization of two subclasses. Virology. 1973 Jul;54(1):270–277. doi: 10.1016/0042-6822(73)90136-0. [DOI] [PubMed] [Google Scholar]
  16. Schnös M., Inman R. B. Position of branch points in replicating lambda DNA. J Mol Biol. 1970 Jul 14;51(1):61–73. doi: 10.1016/0022-2836(70)90270-6. [DOI] [PubMed] [Google Scholar]
  17. Tomizawa J. I., Ohmori H., Bird R. E. Origin of replication of colicin E1 plasmid DNA. Proc Natl Acad Sci U S A. 1977 May;74(5):1865–1869. doi: 10.1073/pnas.74.5.1865. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Tomizawa J., Selzer G. Initiation of DNA synthesis in Escherichia coli. Annu Rev Biochem. 1979;48:999–1034. doi: 10.1146/annurev.bi.48.070179.005031. [DOI] [PubMed] [Google Scholar]
  19. Tsurimoto T., Matsubara K. Purified bacteriophage lambda O protein binds to four repeating sequences at the lambda replication origin. Nucleic Acids Res. 1981 Apr 24;9(8):1789–1799. doi: 10.1093/nar/9.8.1789. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Tsurimoto T., Matsubara K. Replication of lambda dv plasmid in vitro promoted by purified lambda O and P proteins. Proc Natl Acad Sci U S A. 1982 Dec;79(24):7639–7643. doi: 10.1073/pnas.79.24.7639. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Wickner S. H. DNA replication proteins of Escherichia coli and phage lambda. Cold Spring Harb Symp Quant Biol. 1979;43(Pt 1):303–310. doi: 10.1101/sqb.1979.043.01.037. [DOI] [PubMed] [Google Scholar]
  22. Zylicz M., LeBowitz J. H., McMacken R., Georgopoulos C. The dnaK protein of Escherichia coli possesses an ATPase and autophosphorylating activity and is essential in an in vitro DNA replication system. Proc Natl Acad Sci U S A. 1983 Nov;80(21):6431–6435. doi: 10.1073/pnas.80.21.6431. [DOI] [PMC free article] [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