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. 1997 Nov 17;16(22):6886–6895. doi: 10.1093/emboj/16.22.6886

The phiX174-type primosome promotes replisome assembly at the site of recombination in bacteriophage Mu transposition.

J M Jones 1, H Nakai 1
PMCID: PMC1170291  PMID: 9362501

Abstract

Initiation of Escherichia coli DNA synthesis primed by homologous recombination is believed to require the phiX174-type primosome, a mobile priming apparatus assembled without the initiator protein DnaA. We show that this primosome plays an essential role in bacteriophage Mu DNA replication by transposition. Upon promoting transfer of Mu ends to target DNA, the Mu transpososome undergoes transition to a pre-replisome that permits initiation of DNA synthesis only in the presence of primosome assembly proteins PriA, DnaT, DnaB and DnaC. These assembly proteins promote the engagement of primase and DNA polymerase III holoenzyme, initiating semi-discontinuous replication preferentially at the Mu left end. The results indicate that these proteins play a crucial role in promoting replisome assembly on a recombination intermediate.

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Selected References

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  1. Asai T., Kogoma T. D-loops and R-loops: alternative mechanisms for the initiation of chromosome replication in Escherichia coli. J Bacteriol. 1994 Apr;176(7):1807–1812. doi: 10.1128/jb.176.7.1807-1812.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Chaconas G., Lavoie B. D., Watson M. A. DNA transposition: jumping gene machine, some assembly required. Curr Biol. 1996 Jul 1;6(7):817–820. doi: 10.1016/s0960-9822(02)00603-6. [DOI] [PubMed] [Google Scholar]
  3. Craigie R., Mizuuchi K. Transposition of Mu DNA: joining of Mu to target DNA can be uncoupled from cleavage at the ends of Mu. Cell. 1987 Nov 6;51(3):493–501. doi: 10.1016/0092-8674(87)90645-3. [DOI] [PubMed] [Google Scholar]
  4. Craigie R., Mizuuchi M., Mizuuchi K. Site-specific recognition of the bacteriophage Mu ends by the Mu A protein. Cell. 1984 Dec;39(2 Pt 1):387–394. doi: 10.1016/0092-8674(84)90017-5. [DOI] [PubMed] [Google Scholar]
  5. Eggleston A. K., West S. C. Exchanging partners: recombination in E. coli. Trends Genet. 1996 Jan;12(1):20–26. doi: 10.1016/0168-9525(96)81384-9. [DOI] [PubMed] [Google Scholar]
  6. Harshey R. M., McKay R., Bukhari A. I. DNA intermediates in transposition of phage Mu. Cell. 1982 Jun;29(2):561–571. doi: 10.1016/0092-8674(82)90172-6. [DOI] [PubMed] [Google Scholar]
  7. Higgins N. P., Moncecchi D., Manlapaz-Ramos P., Olivera B. M. Bacteriophage Mu DNA replication in vitro. J Biol Chem. 1983 Apr 10;258(7):4293–4297. [PubMed] [Google Scholar]
  8. Kaguni J. M., Kornberg A. Replication initiated at the origin (oriC) of the E. coli chromosome reconstituted with purified enzymes. Cell. 1984 Aug;38(1):183–190. doi: 10.1016/0092-8674(84)90539-7. [DOI] [PubMed] [Google Scholar]
  9. Kogoma T., Cadwell G. W., Barnard K. G., Asai T. The DNA replication priming protein, PriA, is required for homologous recombination and double-strand break repair. J Bacteriol. 1996 Mar;178(5):1258–1264. doi: 10.1128/jb.178.5.1258-1264.1996. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Kogoma T. Recombination by replication. Cell. 1996 May 31;85(5):625–627. doi: 10.1016/s0092-8674(00)81229-5. [DOI] [PubMed] [Google Scholar]
  11. Kruklitis R., Nakai H. Participation of the bacteriophage Mu A protein and host factors in the initiation of Mu DNA synthesis in vitro. J Biol Chem. 1994 Jun 10;269(23):16469–16477. [PubMed] [Google Scholar]
  12. Kruklitis R., Welty D. J., Nakai H. ClpX protein of Escherichia coli activates bacteriophage Mu transposase in the strand transfer complex for initiation of Mu DNA synthesis. EMBO J. 1996 Feb 15;15(4):935–944. [PMC free article] [PubMed] [Google Scholar]
  13. Kuo C. F., Zou A. H., Jayaram M., Getzoff E., Harshey R. DNA-protein complexes during attachment-site synapsis in Mu DNA transposition. EMBO J. 1991 Jun;10(6):1585–1591. doi: 10.1002/j.1460-2075.1991.tb07679.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Lavoie B. D., Chaconas G. Transposition of phage Mu DNA. Curr Top Microbiol Immunol. 1996;204:83–102. doi: 10.1007/978-3-642-79795-8_4. [DOI] [PubMed] [Google Scholar]
  15. Lavoie B. D., Chan B. S., Allison R. G., Chaconas G. Structural aspects of a higher order nucleoprotein complex: induction of an altered DNA structure at the Mu-host junction of the Mu type 1 transpososome. EMBO J. 1991 Oct;10(10):3051–3059. doi: 10.1002/j.1460-2075.1991.tb07856.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Leach D., Symonds N. The isolation and characterisation of a plaque-forming derivative of bacteriophage Mu carrying a fragment of Tn3 conferring ampicillin resistance. Mol Gen Genet. 1979 May 4;172(2):179–184. doi: 10.1007/BF00268280. [DOI] [PubMed] [Google Scholar]
  17. Lee E. H., Kornberg A. Replication deficiencies in priA mutants of Escherichia coli lacking the primosomal replication n' protein. Proc Natl Acad Sci U S A. 1991 Apr 15;88(8):3029–3032. doi: 10.1073/pnas.88.8.3029. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Lee E. H., Masai H., Allen G. C., Jr, Kornberg A. The priA gene encoding the primosomal replicative n' protein of Escherichia coli. Proc Natl Acad Sci U S A. 1990 Jun;87(12):4620–4624. doi: 10.1073/pnas.87.12.4620. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Levchenko I., Luo L., Baker T. A. Disassembly of the Mu transposase tetramer by the ClpX chaperone. Genes Dev. 1995 Oct 1;9(19):2399–2408. doi: 10.1101/gad.9.19.2399. [DOI] [PubMed] [Google Scholar]
  20. Liu J., Nurse P., Marians K. J. The ordered assembly of the phiX174-type primosome. III. PriB facilitates complex formation between PriA and DnaT. J Biol Chem. 1996 Jun 28;271(26):15656–15661. doi: 10.1074/jbc.271.26.15656. [DOI] [PubMed] [Google Scholar]
  21. Maki H., Maki S., Kornberg A. DNA Polymerase III holoenzyme of Escherichia coli. IV. The holoenzyme is an asymmetric dimer with twin active sites. J Biol Chem. 1988 May 15;263(14):6570–6578. [PubMed] [Google Scholar]
  22. Marians K. J. Phi X174-type primosomal proteins: purification and assay. Methods Enzymol. 1995;262:507–521. doi: 10.1016/0076-6879(95)62042-7. [DOI] [PubMed] [Google Scholar]
  23. Masai H., Arai K. Operon structure of dnaT and dnaC genes essential for normal and stable DNA replication of Escherichia coli chromosome. J Biol Chem. 1988 Oct 15;263(29):15083–15093. [PubMed] [Google Scholar]
  24. Masai H., Asai T., Kubota Y., Arai K., Kogoma T. Escherichia coli PriA protein is essential for inducible and constitutive stable DNA replication. EMBO J. 1994 Nov 15;13(22):5338–5345. doi: 10.1002/j.1460-2075.1994.tb06868.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Masukata H., Dasgupta S., Tomizawa J. Transcriptional activation of ColE1 DNA synthesis by displacement of the nontranscribed strand. Cell. 1987 Dec 24;51(6):1123–1130. doi: 10.1016/0092-8674(87)90598-8. [DOI] [PubMed] [Google Scholar]
  26. McBeth D. L., Taylor A. L. Growth of bacteriophage Mu in Escherichia coli dnaA mutants. J Virol. 1982 Nov;44(2):555–564. doi: 10.1128/jvi.44.2.555-564.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. McGlynn P., Al-Deib A. A., Liu J., Marians K. J., Lloyd R. G. The DNA replication protein PriA and the recombination protein RecG bind D-loops. J Mol Biol. 1997 Jul 11;270(2):212–221. doi: 10.1006/jmbi.1997.1120. [DOI] [PubMed] [Google Scholar]
  28. Minden J. S., Marians K. J. Replication of pBR322 DNA in vitro with purified proteins. Requirement for topoisomerase I in the maintenance of template specificity. J Biol Chem. 1985 Aug 5;260(16):9316–9325. [PubMed] [Google Scholar]
  29. Mizuuchi K. In vitro transposition of bacteriophage Mu: a biochemical approach to a novel replication reaction. Cell. 1983 Dec;35(3 Pt 2):785–794. doi: 10.1016/0092-8674(83)90111-3. [DOI] [PubMed] [Google Scholar]
  30. Mizuuchi K. Transpositional recombination: mechanistic insights from studies of mu and other elements. Annu Rev Biochem. 1992;61:1011–1051. doi: 10.1146/annurev.bi.61.070192.005051. [DOI] [PubMed] [Google Scholar]
  31. Mizuuchi M., Baker T. A., Mizuuchi K. Assembly of the active form of the transposase-Mu DNA complex: a critical control point in Mu transposition. Cell. 1992 Jul 24;70(2):303–311. doi: 10.1016/0092-8674(92)90104-k. [DOI] [PubMed] [Google Scholar]
  32. Nakai H., Kruklitis R. Disassembly of the bacteriophage Mu transposase for the initiation of Mu DNA replication. J Biol Chem. 1995 Aug 18;270(33):19591–19598. doi: 10.1074/jbc.270.33.19591. [DOI] [PubMed] [Google Scholar]
  33. Ng J. Y., Marians K. J. The ordered assembly of the phiX174-type primosome. I. Isolation and identification of intermediate protein-DNA complexes. J Biol Chem. 1996 Jun 28;271(26):15642–15648. doi: 10.1074/jbc.271.26.15642. [DOI] [PubMed] [Google Scholar]
  34. Ng J. Y., Marians K. J. The ordered assembly of the phiX174-type primosome. II. Preservation of primosome composition from assembly through replication. J Biol Chem. 1996 Jun 28;271(26):15649–15655. doi: 10.1074/jbc.271.26.15649. [DOI] [PubMed] [Google Scholar]
  35. Nurse P., Zavitz K. H., Marians K. J. Inactivation of the Escherichia coli priA DNA replication protein induces the SOS response. J Bacteriol. 1991 Nov;173(21):6686–6693. doi: 10.1128/jb.173.21.6686-6693.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Resibois A., Pato M., Higgins P., Toussaint A. Replication of bacteriophage mu and its mini-mu derivatives. Adv Exp Med Biol. 1984;179:69–76. doi: 10.1007/978-1-4684-8730-5_7. [DOI] [PubMed] [Google Scholar]
  37. Résibois A., Colet M., Toussaint A. Localisation of mini-Mu in its replication intermediates. EMBO J. 1982;1(8):965–969. doi: 10.1002/j.1460-2075.1982.tb01279.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Résibois A., Toussaint A., Colet M. DNA structures induced by mini-Mu replication. Virology. 1982 Mar;117(2):329–340. doi: 10.1016/0042-6822(82)90473-1. [DOI] [PubMed] [Google Scholar]
  39. Shlomai J., Kornberg A. An Escherichia coli replication protein that recognizes a unique sequence within a hairpin region in phi X174 DNA. Proc Natl Acad Sci U S A. 1980 Feb;77(2):799–803. doi: 10.1073/pnas.77.2.799. [DOI] [PMC free article] [PubMed] [Google Scholar]
  40. Stuitje A. R., Weisbeek P. J., Meijer M. Initiation signals for complementary strand DNA synthesis in the region of the replication origin of the Escherichia coli chromosome. Nucleic Acids Res. 1984 Apr 11;12(7):3321–3332. doi: 10.1093/nar/12.7.3321. [DOI] [PMC free article] [PubMed] [Google Scholar]
  41. Surette M. G., Buch S. J., Chaconas G. Transpososomes: stable protein-DNA complexes involved in the in vitro transposition of bacteriophage Mu DNA. Cell. 1987 Apr 24;49(2):253–262. doi: 10.1016/0092-8674(87)90566-6. [DOI] [PubMed] [Google Scholar]
  42. Tougu K., Peng H., Marians K. J. Identification of a domain of Escherichia coli primase required for functional interaction with the DnaB helicase at the replication fork. J Biol Chem. 1994 Feb 11;269(6):4675–4682. [PubMed] [Google Scholar]
  43. Wickner S., Hurwitz J. Association of phiX174 DNA-dependent ATPase activity with an Escherichia coli protein, replication factor Y, required for in vitro synthesis of phiX174 DNA. Proc Natl Acad Sci U S A. 1975 Sep;72(9):3342–3346. doi: 10.1073/pnas.72.9.3342. [DOI] [PMC free article] [PubMed] [Google Scholar]
  44. Wu C. A., Zechner E. L., Marians K. J. Coordinated leading- and lagging-strand synthesis at the Escherichia coli DNA replication fork. I. Multiple effectors act to modulate Okazaki fragment size. J Biol Chem. 1992 Feb 25;267(6):4030–4044. [PubMed] [Google Scholar]
  45. Yuzhakov A., Turner J., O'Donnell M. Replisome assembly reveals the basis for asymmetric function in leading and lagging strand replication. Cell. 1996 Sep 20;86(6):877–886. doi: 10.1016/s0092-8674(00)80163-4. [DOI] [PubMed] [Google Scholar]
  46. Zipursky S. L., Marians K. J. Identification of two Escherichia coli factor Y effector sites near the origins of replication of the plasmids (ColE1 and pBR322. Proc Natl Acad Sci U S A. 1980 Nov;77(11):6521–6525. doi: 10.1073/pnas.77.11.6521. [DOI] [PMC free article] [PubMed] [Google Scholar]

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