Abstract
We have shown previously that in amino acid-starved, relaxed (rel −) mutants of Escherichia coli replication of the λ plasmid occurs via the λO-containing replication complex (RC) that was assembled prior to the onset of amino acid starvation and is inherited by one of the two daughter plasmid circles in each replication cycle. This replication is regulated neither by binding of the λO initiator to oriλ, nor by the λCro-mediated repression. Here we show that it is dependent on both RNA polymerase and DnaA functions, which is consistent with our recent finding that transcriptional activation of oriλ is under the control of DnaA. In the system studied, DnaA-regulated transcriptional activation of oriλ seems to be the only rate-limiting process. The λ plasmid replication mediated by the inherited RC appeared to be independent of the functions of λP and DnaJ required in RC assembly. In vitro experiments performed by others suggest that DnaJ first binds to the oriλ-bound λO-λO-DnaB pre-primosome and subsequently λP complexed with DnaJ is preferentially recognized by DnaK-GrpE; chaperone-mediated rearrangement of this structure relieves DnaB helicase of λP inhibition. Recently we proposed that this process is directly coupled to the insertion of the pre-primosome between DNA strands transiently separated by transcription. This last-mentioned process may be required in λ plasmid replication mediated by the inherited RC, which appeared in turn to be dependent on DnaK and GrpE functions.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Alfano C, McMacken R (1988) The role of template superhelicity in the initiation of bacteriophage λ DNA replication. Nucleic Acids Res 16:9611–9630
Appleyard RK (1954) Segregation of new lysogenic types during growth of a doubly lysogenic strain derived from Escherichia coli K12. Genetics 39:440–452
Bochner BR, Huang HC, Schieven GL, Ames B (1980) Positive selection for loss of tetracycline resistance. J Bacteriol 143:926–933
Boyd AC, Sherratt DJ (1995) The pCLIP plasmids: versatile cloning vectors based on the bacteriophage λ origin of replication. Gene 153:57–62
Fiil N, Friesen JD (1968) Isolation of relaxed mutants of Escherichia coli. J Bacteriol 95:729–731
Furth ME, Wickner SE (1983) Lambda DNA replication. In: Hendrix RW, Roberts JW, Stahl FW, Weisberg RA (eds) Lambda II. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, New York, pp 145–173
Hansen FG, von Meyenburg K (1979) Characterization of the dnaA, gyrB and other genes in the dnaA region of the Escherichia coli chromosome on specialized transducing phages λtna. Mol Gen Genet 175:135–144
Herman A, Węgrzyn A, Węgrzyn G (1994a) Regulation of replication of plasmid pBR322 in amino acid-starved Escherichia coli strains. Mol Gen Genet 243:374–378
Herman A, Węgrzyn A, Węgrzyn G (1994b) Differential replication of plasmids during stringent and relaxed response of Escherichia coli. Plasmid 32:89–94
Klinkert J, Klein A (1978) Roles of bacteriophage lambda gene products O and P during early and late phases of infection cycle. J Virol 25:730–737
Kornberg A, Baker TA (1992) DNA replication. WH Freeman, New York
Kur J, Górska I, Taylor K (1987) Escherichia coli dnaA initiation function is required for replication of plasmids derived from coliphage lambda. J Mol Biol 198:203–210
Mensa-Wilmot K, Carroll K, McMacken R (1989) Transcriptional activation of bacteriophage λ DNA replication in vitro: regulatory role of histone-like protein HU of Escherichia coli. EMBO J 8:2393–2402
Osipiuk J, Georgopoulos C, Zylicz M (1993) Initiation of 7 DNA replication: The Escherichia coli small heat shock proteins, DnaJ and GrpE, increase DnaK's affinity for the λP protein. J Biol Chem 268:4821–4827
Pawłowicz A, Węgrzyn G, Taylor K (1993) Effect of coliphage λP gene mutations on the stability of the λO protein, the initiator of λ DNA replication. Acta Biochim Polon 40:29–31
Sambrook J, Fritsch EF, Maniatis T (1989) Molecular cloning: a Laboratory manual. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, New York
Silhavy TJ, Berman ML, Enquist LW (1984) Experiments with gene fusions. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, New York
Szalewska A, Węgrzyn G, Taylor K (1994) Neither absence nor excess of 70 digesting ClpXP protease affects 7 plasmid or phage replication in Escherichia coli. Mol Microbiol 13:469–474
Szalewska-Pałasz A, Węgrzyn A, Herman A, Węgrzyn G (1994) The mechanism of the stringent control of λ plasmid DNA replication. EMBO J 13:5779–5785
Taylor K, Węgrzyn G (1995) Replication of coliphage λ DNA. FEMS Microbiol Rev in press
Węgrzyn A, Węgrzyn G, Taylor K (1995) Protection of coliphage λO initiator protein from proteolysis in the assembly of the replication complex in vivo. Virology 207:179–184
Węgrzyn G, Taylor K (1992) Inheritance of the replication complex by one of two daughter copies during λ plasmid replication in Escherichia coli. J Mol Biol 226:681–688
Węgrzyn G, Pawłowicz A, Taylor K (1992) Stability of coliphage DNA replication initiator, the λO protein. J Mol Biol 226:675–680
Węgrzyn G, Neubauer P, Krueger S, Hecker M, Taylor K (1991) Stringent control of replication of plasmids derived from coliphage λ. Mol Gen Genet 225:94–98
Węgrzyn G, Szalewska-Pałasz A, Węgrzyn A, Obuchowski M, Taylor K (1995a) Transcriptional activation of the origin of coliphage lambda DNA replication is regulated by the host DnaA initiator function. Gene 154:47–50
Węgrzyn G, Węgrzyn A, Konieczny I, Bielawski K, Konopa G, Obuchowski M, Helinski DR, Taylor K (1995b)Involvement of the host initiator function dnaA in the replication of coliphage λ. J. Genetics 139: in press
Xiao H, Kalman M, Ikehara K, Zemel S, Glaser G, Cashel M (1991) Residual guanosine 3′,5′-bipyrophosphate synthetic activity of relA null mutants can be eliminated by spoT null mutations. J Biol Chem 266:5980–5990
Zylicz M (1993) The Escherichia coli chaperones involved in DNA replication. Phil Trans R Soc Lond 339:271–278
Author information
Authors and Affiliations
Additional information
Communicated by R. Devoret
Rights and permissions
About this article
Cite this article
Węgrzyn, A., Węgrzyn, G. & Taylor, K. Plasmid and host functions required for λ plasmid replication carried out by the inherited replication complex. Molec. Gen. Genet. 247, 501–508 (1995). https://doi.org/10.1007/BF00293153
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF00293153