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. 1989 Apr;171(4):2056–2065. doi: 10.1128/jb.171.4.2056-2065.1989

Involvement of integration host factor (IHF) in maintenance of plasmid pSC101 in Escherichia coli: characterization of pSC101 mutants that replicate in the absence of IHF.

D P Biek 1, S N Cohen 1
PMCID: PMC209857  PMID: 2539358

Abstract

Escherichia coli mutants defective in the stable maintenance of plasmid pSC101 have been isolated following Tn10 insertion mutagenesis. One class of mutations affecting pSC101 replication was located in the genes himA and himD (hip), which encode the two subunits of integration host factor (IHF), a small histonelike DNA-binding protein that has multiple cellular functions. Mutants of pSC101 that could replicate in the absence of IHF were isolated and characterized; four independent mutational alterations were found to affect the third codon of the pSC101 rep gene, resulting in the replacement of glutamic acid by lysine. The compensating alteration appears to function by altering the activity of the pSC101 rep protein in him mutants.

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Selected References

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  1. Abeles A. L., Friedman S. A., Austin S. J. Partition of unit-copy miniplasmids to daughter cells. III. The DNA sequence and functional organization of the P1 partition region. J Mol Biol. 1985 Sep 20;185(2):261–272. doi: 10.1016/0022-2836(85)90402-4. [DOI] [PubMed] [Google Scholar]
  2. Amundsen S. K., Taylor A. F., Chaudhury A. M., Smith G. R. recD: the gene for an essential third subunit of exonuclease V. Proc Natl Acad Sci U S A. 1986 Aug;83(15):5558–5562. doi: 10.1073/pnas.83.15.5558. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Armstrong K. A., Acosta R., Ledner E., Machida Y., Pancotto M., McCormick M., Ohtsubo H., Ohtsubo E. A 37 X 10(3) molecular weight plasmid-encoded protein is required for replication and copy number control in the plasmid pSC101 and its temperature-sensitive derivative pHS1. J Mol Biol. 1984 May 25;175(3):331–348. doi: 10.1016/0022-2836(84)90352-8. [DOI] [PubMed] [Google Scholar]
  4. Bachmann B. J. Linkage map of Escherichia coli K-12, edition 7. Microbiol Rev. 1983 Jun;47(2):180–230. doi: 10.1128/mr.47.2.180-230.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Bernardi A., Bernardi F. Complete sequence of pSC101. Nucleic Acids Res. 1984 Dec 21;12(24):9415–9426. doi: 10.1093/nar/12.24.9415. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Biek D. P., Cohen S. N. Identification and characterization of recD, a gene affecting plasmid maintenance and recombination in Escherichia coli. J Bacteriol. 1986 Aug;167(2):594–603. doi: 10.1128/jb.167.2.594-603.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Biek D. P., Cohen S. N. Involvement of integration host factor (IHF) in maintenance of plasmid pSC101 in Escherichia coli: mutations in the topA gene allow pSC101 replication in the absence of IHF. J Bacteriol. 1989 Apr;171(4):2066–2074. doi: 10.1128/jb.171.4.2066-2074.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Cohen S. N., Chang A. C. Revised interpretation of the origin of the pSC101 plasmid. J Bacteriol. 1977 Nov;132(2):734–737. doi: 10.1128/jb.132.2.734-737.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Doherty M. J., Morrison P. T., Kolodner R. Genetic recombination of bacterial plasmid DNA. Physical and genetic analysis of the products of plasmid recombination in Escherichia coli. J Mol Biol. 1983 Jul 5;167(3):539–560. doi: 10.1016/s0022-2836(83)80097-7. [DOI] [PubMed] [Google Scholar]
  10. Dorman C. J., Higgins C. F. Fimbrial phase variation in Escherichia coli: dependence on integration host factor and homologies with other site-specific recombinases. J Bacteriol. 1987 Aug;169(8):3840–3843. doi: 10.1128/jb.169.8.3840-3843.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Drlica K., Rouviere-Yaniv J. Histonelike proteins of bacteria. Microbiol Rev. 1987 Sep;51(3):301–319. doi: 10.1128/mr.51.3.301-319.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Ely S., Wright A. Maintenance of plasmid pSC101 in Escherichia coli requires the host primase. J Bacteriol. 1985 Oct;164(1):484–486. doi: 10.1128/jb.164.1.484-486.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Feiss M., Fogarty S., Christiansen S. Bacteriophage lambda DNA packaging: a mutant terminase that is independent of integration host factor. Mol Gen Genet. 1988 Apr;212(1):142–148. doi: 10.1007/BF00322457. [DOI] [PubMed] [Google Scholar]
  14. Feiss M., Frackman S., Sippy J. Essential interaction between lambdoid phage 21 terminase and the Escherichia coli integrative host factor. J Mol Biol. 1985 May 25;183(2):239–246. doi: 10.1016/0022-2836(85)90216-5. [DOI] [PubMed] [Google Scholar]
  15. Flamm E. L., Weisberg R. A. Primary structure of the hip gene of Escherichia coli and of its product, the beta subunit of integration host factor. J Mol Biol. 1985 May 25;183(2):117–128. doi: 10.1016/0022-2836(85)90206-2. [DOI] [PubMed] [Google Scholar]
  16. Frey J., Chandler M., Caro L. The effects of an Escherichia coli dnaAts mutation on the replication of the plasmids colE1 pSC101, R100.1 and RTF-TC. Mol Gen Genet. 1979 Jul 13;174(2):117–126. doi: 10.1007/BF00268349. [DOI] [PubMed] [Google Scholar]
  17. Fuller R. S., Funnell B. E., Kornberg A. The dnaA protein complex with the E. coli chromosomal replication origin (oriC) and other DNA sites. Cell. 1984 Oct;38(3):889–900. doi: 10.1016/0092-8674(84)90284-8. [DOI] [PubMed] [Google Scholar]
  18. Funnell B. E. Participation of Escherichia coli integration host factor in the P1 plasmid partition system. Proc Natl Acad Sci U S A. 1988 Sep;85(18):6657–6661. doi: 10.1073/pnas.85.18.6657. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Gamas P., Burger A. C., Churchward G., Caro L., Galas D., Chandler M. Replication of pSC101: effects of mutations in the E. coli DNA binding protein IHF. Mol Gen Genet. 1986 Jul;204(1):85–89. doi: 10.1007/BF00330192. [DOI] [PubMed] [Google Scholar]
  20. Gerdes K., Molin S. Partitioning of plasmid R1. Structural and functional analysis of the parA locus. J Mol Biol. 1986 Aug 5;190(3):269–279. doi: 10.1016/0022-2836(86)90001-x. [DOI] [PubMed] [Google Scholar]
  21. Goosen N., van de Putte P. Regulation of Mu transposition. I. Localization of the presumed recognition sites for HimD and Ner functions controlling bacteriophage Mu transcription. Gene. 1984 Oct;30(1-3):41–46. doi: 10.1016/0378-1119(84)90103-3. [DOI] [PubMed] [Google Scholar]
  22. Granston A. E., Alessi D. M., Eades L. J., Friedman D. I. A point mutation in the Nul gene of bacteriophage lambda facilitates phage growth in Escherichia coli with himA and gyrB mutations. Mol Gen Genet. 1988 Apr;212(1):149–156. doi: 10.1007/BF00322458. [DOI] [PubMed] [Google Scholar]
  23. Greenstein D., Zinder N. D., Horiuchi K. Integration host factor interacts with the DNA replication enhancer of filamentous phage f1. Proc Natl Acad Sci U S A. 1988 Sep;85(17):6262–6266. doi: 10.1073/pnas.85.17.6262. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Gustafsson P., Wolf-Watz H., Lind L., Johansson K. E., Nordström K. Binding between the par region of plasmids R1 and pSC101 and the outer membrane fraction of the host bacteria. EMBO J. 1983;2(1):27–32. doi: 10.1002/j.1460-2075.1983.tb01375.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Guyer M. S., Reed R. R., Steitz J. A., Low K. B. Identification of a sex-factor-affinity site in E. coli as gamma delta. Cold Spring Harb Symp Quant Biol. 1981;45(Pt 1):135–140. doi: 10.1101/sqb.1981.045.01.022. [DOI] [PubMed] [Google Scholar]
  26. Hashimoto T., Sekiguchi M. Isolation of temperature-sensitive mutants of R plasmid by in vitro mutagenesis with hydroxylamine. J Bacteriol. 1976 Sep;127(3):1561–1563. doi: 10.1128/jb.127.3.1561-1563.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Hasunuma K., Sekiguchi M. Effect of dna mutations on the replication of plasmid pSC101 in Escherichia coli K-12. J Bacteriol. 1979 Mar;137(3):1095–1099. doi: 10.1128/jb.137.3.1095-1099.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Hirota Y., Mordoh J., Jacob F. On the process of cellular division in Escherichia coli. 3. Thermosensitive mutants of Escherichia coli altered in the process of DNA initiation. J Mol Biol. 1970 Nov 14;53(3):369–387. doi: 10.1016/0022-2836(70)90072-0. [DOI] [PubMed] [Google Scholar]
  29. James A. A., Morrison P. T., Kolodner R. Genetic recombination of bacterial plasmid DNA. Analysis of the effect of recombination-deficient mutations on plasmid recombination. J Mol Biol. 1982 Sep 25;160(3):411–430. doi: 10.1016/0022-2836(82)90305-9. [DOI] [PubMed] [Google Scholar]
  30. Kikuchi A., Flamm E., Weisberg R. A. An Escherichia coli mutant unable to support site-specific recombination of bacteriophage lambda. J Mol Biol. 1985 May 25;183(2):129–140. doi: 10.1016/0022-2836(85)90207-4. [DOI] [PubMed] [Google Scholar]
  31. Kline B. C., Trawick J. Identification and characterization of a second copy number control gene in mini-F plasmids. Mol Gen Genet. 1983;192(3):408–415. doi: 10.1007/BF00392183. [DOI] [PubMed] [Google Scholar]
  32. Krause H. M., Higgins N. P. Positive and negative regulation of the Mu operator by Mu repressor and Escherichia coli integration host factor. J Biol Chem. 1986 Mar 15;261(8):3744–3752. [PubMed] [Google Scholar]
  33. Lange-Gustafson B. J., Nash H. A. Purification and properties of Int-h, a variant protein involved in site-specific recombination of bacteriophage lambda. J Biol Chem. 1984 Oct 25;259(20):12724–12732. [PubMed] [Google Scholar]
  34. Linder P., Churchward G., Caro L. Plasmid pSC101 replication mutants generated by insertion of the transposon Tn1000. J Mol Biol. 1983 Oct 25;170(2):287–303. doi: 10.1016/s0022-2836(83)80149-1. [DOI] [PubMed] [Google Scholar]
  35. Meacock P. A., Cohen S. N. Partitioning of bacterial plasmids during cell division: a cis-acting locus that accomplishes stable plasmid inheritance. Cell. 1980 Jun;20(2):529–542. doi: 10.1016/0092-8674(80)90639-x. [DOI] [PubMed] [Google Scholar]
  36. Miller C. A., Tucker W. T., Meacock P. A., Gustafsson P., Cohen S. N. Nucleotide sequence of the partition locus of Escherichia coli plasmid pSC101. Gene. 1983 Oct;24(2-3):309–315. doi: 10.1016/0378-1119(83)90091-4. [DOI] [PubMed] [Google Scholar]
  37. Miller H. I., Friedman D. I. An E. coli gene product required for lambda site-specific recombination. Cell. 1980 Jul;20(3):711–719. doi: 10.1016/0092-8674(80)90317-7. [DOI] [PubMed] [Google Scholar]
  38. Miller H. I., Kikuchi A., Nash H. A., Weisberg R. A., Friedman D. I. Site-specific recombination of bacteriophage lambda: the role of host gene products. Cold Spring Harb Symp Quant Biol. 1979;43(Pt 2):1121–1126. doi: 10.1101/sqb.1979.043.01.125. [DOI] [PubMed] [Google Scholar]
  39. Miller H. I., Mozola M. A., Friedman D. I. int-h: An int mutation of phage lambda that enhances site-specific recombination. Cell. 1980 Jul;20(3):721–729. doi: 10.1016/0092-8674(80)90318-9. [DOI] [PubMed] [Google Scholar]
  40. Miller H. I., Nash H. A. Direct role of the himA gene product in phage lambda integration. Nature. 1981 Apr 9;290(5806):523–526. doi: 10.1038/290523a0. [DOI] [PubMed] [Google Scholar]
  41. Miller H. I. Primary structure of the himA gene of Escherichia coli: homology with DNA-binding protein HU and association with the phenylalanyl-tRNA synthetase operon. Cold Spring Harb Symp Quant Biol. 1984;49:691–698. doi: 10.1101/sqb.1984.049.01.078. [DOI] [PubMed] [Google Scholar]
  42. Mori H., Kondo A., Ohshima A., Ogura T., Hiraga S. Structure and function of the F plasmid genes essential for partitioning. J Mol Biol. 1986 Nov 5;192(1):1–15. doi: 10.1016/0022-2836(86)90459-6. [DOI] [PubMed] [Google Scholar]
  43. Morisato D., Kleckner N. Tn10 transposition and circle formation in vitro. Cell. 1987 Oct 9;51(1):101–111. doi: 10.1016/0092-8674(87)90014-6. [DOI] [PubMed] [Google Scholar]
  44. Nordström K., Molin S., Aagaard-Hansen H. Partitioning of plasmid R1 in Escherichia coli. I. Kinetics of loss of plasmid derivatives deleted of the par region. Plasmid. 1980 Sep;4(2):215–227. doi: 10.1016/0147-619x(80)90011-6. [DOI] [PubMed] [Google Scholar]
  45. Norrander J., Kempe T., Messing J. Construction of improved M13 vectors using oligodeoxynucleotide-directed mutagenesis. Gene. 1983 Dec;26(1):101–106. doi: 10.1016/0378-1119(83)90040-9. [DOI] [PubMed] [Google Scholar]
  46. Prentki P., Chandler M., Galas D. J. Escherichia coli integration host factor bends the DNA at the ends of IS1 and in an insertion hotspot with multiple IHF binding sites. EMBO J. 1987 Aug;6(8):2479–2487. doi: 10.1002/j.1460-2075.1987.tb02529.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  47. Reiss B., Sprengel R., Schaller H. Protein fusions with the kanamycin resistance gene from transposon Tn5. EMBO J. 1984 Dec 20;3(13):3317–3322. doi: 10.1002/j.1460-2075.1984.tb02297.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  48. Richet E., Abcarian P., Nash H. A. The interaction of recombination proteins with supercoiled DNA: defining the role of supercoiling in lambda integrative recombination. Cell. 1986 Sep 26;46(7):1011–1021. doi: 10.1016/0092-8674(86)90700-2. [DOI] [PubMed] [Google Scholar]
  49. Robertson C. A., Nash H. A. Bending of the bacteriophage lambda attachment site by Escherichia coli integration host factor. J Biol Chem. 1988 Mar 15;263(8):3554–3557. [PubMed] [Google Scholar]
  50. Sanger F., Nicklen S., Coulson A. R. DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci U S A. 1977 Dec;74(12):5463–5467. doi: 10.1073/pnas.74.12.5463. [DOI] [PMC free article] [PubMed] [Google Scholar]
  51. Stenzel T. T., Patel P., Bastia D. The integration host factor of Escherichia coli binds to bent DNA at the origin of replication of the plasmid pSC101. Cell. 1987 Jun 5;49(5):709–717. doi: 10.1016/0092-8674(87)90547-2. [DOI] [PubMed] [Google Scholar]
  52. Tautz D., Renz M. An optimized freeze-squeeze method for the recovery of DNA fragments from agarose gels. Anal Biochem. 1983 Jul 1;132(1):14–19. doi: 10.1016/0003-2697(83)90419-0. [DOI] [PubMed] [Google Scholar]
  53. Thompson J. F., Landy A. Empirical estimation of protein-induced DNA bending angles: applications to lambda site-specific recombination complexes. Nucleic Acids Res. 1988 Oct 25;16(20):9687–9705. doi: 10.1093/nar/16.20.9687. [DOI] [PMC free article] [PubMed] [Google Scholar]
  54. Thompson J. F., de Vargas L. M., Skinner S. E., Landy A. Protein-protein interactions in a higher-order structure direct lambda site-specific recombination. J Mol Biol. 1987 Jun 5;195(3):481–493. doi: 10.1016/0022-2836(87)90177-x. [DOI] [PubMed] [Google Scholar]
  55. Tucker W. T., Miller C. A., Cohen S. N. Structural and functional analysis of the par region of the pSC 10 1 plasmid. Cell. 1984 Aug;38(1):191–201. doi: 10.1016/0092-8674(84)90540-3. [DOI] [PubMed] [Google Scholar]
  56. Uhlin B. E., Nordström K. R plasmid gene dosage effects in Escherichia coli K-12: copy mutants of the R plasmic R1drd-19. Plasmid. 1977 Nov;1(1):1–7. doi: 10.1016/0147-619x(77)90003-8. [DOI] [PubMed] [Google Scholar]
  57. Vocke C., Bastia D. Primary structure of the essential replicon of the plasmid pSC101. Proc Natl Acad Sci U S A. 1983 Nov;80(21):6557–6561. doi: 10.1073/pnas.80.21.6557. [DOI] [PMC free article] [PubMed] [Google Scholar]
  58. Wahle E., Kornberg A. The partition locus of plasmid pSC101 is a specific binding site for DNA gyrase. EMBO J. 1988 Jun;7(6):1889–1895. doi: 10.1002/j.1460-2075.1988.tb03022.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  59. Yamaguchi K., Masamune Y. Autogenous regulation of synthesis of the replication protein in plasmid pSC101. Mol Gen Genet. 1985;200(3):362–367. doi: 10.1007/BF00425718. [DOI] [PubMed] [Google Scholar]

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