Structural and Functional Analysis of the Origin of Conjugal Transfer of the Broad-host-range IncW Plasmid R388 and Comparison with the Related IncN Plasmid R46

Overview

Journal Mol Gen Genet

Specialties Genetics
Molecular Biology

Date 1991 May 1

PMID 2038309

Citations 23

Authors

M Llosa

S Bolland

F de la Cruz

Affiliations

Soon will be listed here.

Abstract

We cloned and sequenced a 402 bp DNA segment containing the origin of conjugal transfer (oriT) of the IncW plasmid R388. Progressive deletions from each end of the sequence were assayed for oriT activity. Stepwise reductions in mobilization frequencies, representing the loss of functional elements, correlated with deletion of structural motifs in the sequence. A sequence of 330 bp of oriT was sufficient for efficient mobilization. The first 86 bp of the sequence contains five tandemly repeated DNA sequences of 11 bp, followed by a 10 bp perfect inverted repeat. Deletion of the first 95 bp reduced the frequency of transfer by a hundred-fold. The sequence between bp 183 and 218 was necessary and sufficient for low frequency mobilization and, thus, it was assumed to contain the nick site. This basis core was cloned as a 60 bp segment (from bp 176-236) that could be mobilized at low frequency. It includes two inverted repeats and a perfect integration host factor (IHF) consensus binding site. A third functionally important segment in oriT was located between bp 260 and 330. The DNA sequence of the oriT of R388 could be aligned with that of the broad-host-range IncN plasmid R46. Moreover, the relative positions of the three inverted repeats are also conserved. Overall sequence similarity was 52%, but was significantly higher in particular regions, which coincided with the functionally important segments mapped by deletion analysis. Conservation of these segments provided independent support for their essential role in oriT function.

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References

McIntire S, Dempsey W . oriT sequence of the antibiotic resistance plasmid R100. J Bacteriol. 1987; 169(8):3829-32. PMC: 212476. DOI: 10.1128/jb.169.8.3829-3832.1987. View

Brown A, Willetts N . A physical and genetic map of the IncN plasmid R46. Plasmid. 1981; 5(2):188-201. DOI: 10.1016/0147-619x(81)90020-2. View

. RecA independent, site-specific recombination between ColE1 or ColK and a miniplasmid they complement for mobilization and relaxation: implications for the mechanism of DNA transfer during mobilization. Plasmid. 1980; 4(1):51-63. DOI: 10.1016/0147-619x(80)90082-7. View

Yamamoto T, Motegi A, Takei T, Okayama H, Sawai T . Plasmid R46 provides a function that promotes recA-independent deletion, fusion and resolution of replicon. Mol Gen Genet. 1984; 193(2):255-62. DOI: 10.1007/BF00330677. View

Derbyshire K, Willetts N . Mobilization of the non-conjugative plasmid RSF1010: a genetic analysis of its origin of transfer. Mol Gen Genet. 1987; 206(1):154-60. DOI: 10.1007/BF00326551. View

Leong J, Lesser C, Youderian P, Susskind M, Landy A . The phi 80 and P22 attachment sites. Primary structure and interaction with Escherichia coli integration host factor. J Biol Chem. 1985; 260(7):4468-77. View

Wang J, Giaever G . Action at a distance along a DNA. Science. 1988; 240(4850):300-4. DOI: 10.1126/science.3281259. View

Goldner A, Graus H, Hogenauer G . The origin of transfer of P307. Plasmid. 1987; 18(1):76-83. DOI: 10.1016/0147-619x(87)90080-1. View

Goto N, Shoji A, Horiuchi S, Nakaya R . Conduction of nonconjugative plasmids by F' lac is not necessarily associated with transposition of the gamma delta sequence. J Bacteriol. 1984; 159(2):590-6. PMC: 215684. DOI: 10.1128/jb.159.2.590-596.1984. View

10.

Friden P, VOELKEL K, Sternglanz R, Freundlich M . Reduced expression of the isoleucine and valine enzymes in integration host factor mutants of Escherichia coli. J Mol Biol. 1984; 172(4):573-9. DOI: 10.1016/s0022-2836(84)80024-8. View

11.

Thompson T, Centola M, Deonier R . Location of the nick at oriT of the F plasmid. J Mol Biol. 1989; 207(3):505-12. DOI: 10.1016/0022-2836(89)90460-9. View

12.

Perez-Martin J, del Solar G, de la Campa A, Espinosa M . Three regions in the DNA of plasmid pLS1 show sequence-directed static bending. Nucleic Acids Res. 1988; 16(19):9113-26. PMC: 338695. DOI: 10.1093/nar/16.19.9113. View

13.

Pansegrau W, Ziegelin G, LANKA E . The origin of conjugative IncP plasmid transfer: interaction with plasmid-encoded products and the nucleotide sequence at the relaxation site. Biochim Biophys Acta. 1988; 951(2-3):365-74. DOI: 10.1016/0167-4781(88)90108-x. View

14.

Warren G, Clark A . Sequence-specific recombination of plasmid ColE1. Proc Natl Acad Sci U S A. 1980; 77(11):6724-8. PMC: 350361. DOI: 10.1073/pnas.77.11.6724. View

15.

Tsai M, Fu Y, Deonier R . Intrinsic bends and integration host factor binding at F plasmid oriT. J Bacteriol. 1990; 172(8):4603-9. PMC: 213294. DOI: 10.1128/jb.172.8.4603-4609.1990. View

16.

de la Cruz F, Grinsted J . Genetic and molecular characterization of Tn21, a multiple resistance transposon from R100.1. J Bacteriol. 1982; 151(1):222-28. PMC: 220230. DOI: 10.1128/jb.151.1.222-228.1982. View

17.

Ziegelin G, Furste J, LANKA E . TraJ protein of plasmid RP4 binds to a 19-base pair invert sequence repetition within the transfer origin. J Biol Chem. 1989; 264(20):11989-94. View

18.

Willetts N, Wilkins B . Processing of plasmid DNA during bacterial conjugation. Microbiol Rev. 1984; 48(1):24-41. PMC: 373001. DOI: 10.1128/mr.48.1.24-41.1984. View

19.

Vieira J, Messing J . The pUC plasmids, an M13mp7-derived system for insertion mutagenesis and sequencing with synthetic universal primers. Gene. 1982; 19(3):259-68. DOI: 10.1016/0378-1119(82)90015-4. View

20.

Ostermann E, Kricek F, Hogenauer G . Cloning the origin of transfer region of the resistance plasmid R1. EMBO J. 1984; 3(8):1731-5. PMC: 557589. DOI: 10.1002/j.1460-2075.1984.tb02039.x. View