Abstract
NBU1 is a 10.3-kbp integrated Bacteroides element that can be induced to excise from the chromosome and can be mobilized to a recipient by trans-acting functions provided by certain Bacteroides conjugative transposons. The NBU1 transfer intermediate is a covalently closed circle, which is presumed to be the form that integrates into the recipient genome. We report here that a 2.4-kbp segment of NBU1 was all that was required for site-specific integration into the chromosome of Bacteroides thetaiotaomicron 5482. This 2.4-kbp region included the joined ends of the NBU1 circular form (attN1) and a single open reading frame, intN1, which encoded the integrase. Previously, we had found that NBU1 integrates preferentially into a single site in B. thetaiotaomicron 5482. We have now shown that the NBU1 target site is located at the 3' end of a Leu-tRNA gene. The NBU1 integrase gene, intN1, was sequenced. The predicted protein had little overall amino acid sequence similarity to any proteins in the databases but had limited carboxy-terminal similarity to the integrases of lambdoid phages and to the integrases of the gram-positive conjugative transposons Tn916 and Tn1545. We also report that the intN1 gene is expressed constitutively.
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- Altschul S. F., Gish W., Miller W., Myers E. W., Lipman D. J. Basic local alignment search tool. J Mol Biol. 1990 Oct 5;215(3):403–410. doi: 10.1016/S0022-2836(05)80360-2. [DOI] [PubMed] [Google Scholar]
- Argos P., Landy A., Abremski K., Egan J. B., Haggard-Ljungquist E., Hoess R. H., Kahn M. L., Kalionis B., Narayana S. V., Pierson L. S., 3rd The integrase family of site-specific recombinases: regional similarities and global diversity. EMBO J. 1986 Feb;5(2):433–440. doi: 10.1002/j.1460-2075.1986.tb04229.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Bedzyk L. A., Shoemaker N. B., Young K. E., Salyers A. A. Insertion and excision of Bacteroides conjugative chromosomal elements. J Bacteriol. 1992 Jan;174(1):166–172. doi: 10.1128/jb.174.1.166-172.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Boccard F., Smokvina T., Pernodet J. L., Friedmann A., Guérineau M. The integrated conjugative plasmid pSAM2 of Streptomyces ambofaciens is related to temperate bacteriophages. EMBO J. 1989 Mar;8(3):973–980. doi: 10.1002/j.1460-2075.1989.tb03460.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Brown D. P., Idler K. B., Katz L. Characterization of the genetic elements required for site-specific integration of plasmid pSE211 in Saccharopolyspora erythraea. J Bacteriol. 1990 Apr;172(4):1877–1888. doi: 10.1128/jb.172.4.1877-1888.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Campbell A. M. Chromosomal insertion sites for phages and plasmids. J Bacteriol. 1992 Dec;174(23):7495–7499. doi: 10.1128/jb.174.23.7495-7499.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Devereux J., Haeberli P., Smithies O. A comprehensive set of sequence analysis programs for the VAX. Nucleic Acids Res. 1984 Jan 11;12(1 Pt 1):387–395. doi: 10.1093/nar/12.1part1.387. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Feldhaus M. J., Hwa V., Cheng Q., Salyers A. A. Use of an Escherichia coli beta-glucuronidase gene as a reporter gene for investigation of Bacteroides promoters. J Bacteriol. 1991 Jul;173(14):4540–4543. doi: 10.1128/jb.173.14.4540-4543.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Kiliç A. O., Vijayakumar M. N., al-Khaldi S. F. Identification and nucleotide sequence analysis of a transfer-related region in the streptococcal conjugative transposon Tn5252. J Bacteriol. 1994 Aug;176(16):5145–5150. doi: 10.1128/jb.176.16.5145-5150.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Komine Y., Adachi T., Inokuchi H., Ozeki H. Genomic organization and physical mapping of the transfer RNA genes in Escherichia coli K12. J Mol Biol. 1990 Apr 20;212(4):579–598. doi: 10.1016/0022-2836(90)90224-A. [DOI] [PubMed] [Google Scholar]
- Li L. Y., Shoemaker N. B., Salyers A. A. Characterization of the mobilization region of a Bacteroides insertion element (NBU1) that is excised and transferred by Bacteroides conjugative transposons. J Bacteriol. 1993 Oct;175(20):6588–6598. doi: 10.1128/jb.175.20.6588-6598.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Li L. Y., Shoemaker N. B., Salyers A. A. Location and characteristics of the transfer region of a Bacteroides conjugative transposon and regulation of transfer genes. J Bacteriol. 1995 Sep;177(17):4992–4999. doi: 10.1128/jb.177.17.4992-4999.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Li L. Y., Shoemaker N. B., Wang G. R., Cole S. P., Hashimoto M. K., Wang J., Salyers A. A. The mobilization regions of two integrated Bacteroides elements, NBU1 and NBU2, have only a single mobilization protein and may be on a cassette. J Bacteriol. 1995 Jul;177(14):3940–3945. doi: 10.1128/jb.177.14.3940-3945.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Poyart-Salmeron C., Trieu-Cuot P., Carlier C., Courvalin P. Molecular characterization of two proteins involved in the excision of the conjugative transposon Tn1545: homologies with other site-specific recombinases. EMBO J. 1989 Aug;8(8):2425–2433. doi: 10.1002/j.1460-2075.1989.tb08373.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Rauch P. J., de Vos W. M. Identification and characterization of genes involved in excision of the Lactococcus lactis conjugative transposon Tn5276. J Bacteriol. 1994 Apr;176(8):2165–2171. doi: 10.1128/jb.176.8.2165-2171.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
- SAITO H., MIURA K. I. PREPARATION OF TRANSFORMING DEOXYRIBONUCLEIC ACID BY PHENOL TREATMENT. Biochim Biophys Acta. 1963 Aug 20;72:619–629. [PubMed] [Google Scholar]
- Shoemaker N. B., Getty C., Guthrie E. P., Salyers A. A. Regions in Bacteroides plasmids pBFTM10 and pB8-51 that allow Escherichia coli-Bacteroides shuttle vectors to be mobilized by IncP plasmids and by a conjugative Bacteroides tetracycline resistance element. J Bacteriol. 1986 Jun;166(3):959–965. doi: 10.1128/jb.166.3.959-965.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Shoemaker N. B., Li L. Y., Salyers A. A. An unusual type of cointegrate formation between a Bacteroides plasmid and the excised circular form of an integrated element (NBU1). Plasmid. 1994 Nov;32(3):312–317. doi: 10.1006/plas.1994.1070. [DOI] [PubMed] [Google Scholar]
- Shoemaker N. B., Salyers A. A. A cryptic 65-kilobase-pair transposonlike element isolated from Bacteroides uniformis has homology with Bacteroides conjugal tetracycline resistance elements. J Bacteriol. 1990 Apr;172(4):1694–1702. doi: 10.1128/jb.172.4.1694-1702.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Shoemaker N. B., Salyers A. A. Tetracycline-dependent appearance of plasmidlike forms in Bacteroides uniformis 0061 mediated by conjugal Bacteroides tetracycline resistance elements. J Bacteriol. 1988 Apr;170(4):1651–1657. doi: 10.1128/jb.170.4.1651-1657.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Shoemaker N. B., Wang G. R., Stevens A. M., Salyers A. A. Excision, transfer, and integration of NBU1, a mobilizable site-selective insertion element. J Bacteriol. 1993 Oct;175(20):6578–6587. doi: 10.1128/jb.175.20.6578-6587.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Smith C. J., Parker A. C. Identification of a circular intermediate in the transfer and transposition of Tn4555, a mobilizable transposon from Bacteroides spp. J Bacteriol. 1993 May;175(9):2682–2691. doi: 10.1128/jb.175.9.2682-2691.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Stevens A. M., Sanders J. M., Shoemaker N. B., Salyers A. A. Genes involved in production of plasmidlike forms by a Bacteroides conjugal chromosomal element share amino acid homology with two-component regulatory systems. J Bacteriol. 1992 May;174(9):2935–2942. doi: 10.1128/jb.174.9.2935-2942.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Stevens A. M., Shoemaker N. B., Salyers A. A. The region of a Bacteroides conjugal chromosomal tetracycline resistance element which is responsible for production of plasmidlike forms from unlinked chromosomal DNA might also be involved in transfer of the element. J Bacteriol. 1990 Aug;172(8):4271–4279. doi: 10.1128/jb.172.8.4271-4279.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Su Y. A., Clewell D. B. Characterization of the left 4 kb of conjugative transposon Tn916: determinants involved in excision. Plasmid. 1993 Nov;30(3):234–250. doi: 10.1006/plas.1993.1055. [DOI] [PubMed] [Google Scholar]
- Valentine P. J., Shoemaker N. B., Salyers A. A. Mobilization of Bacteroides plasmids by Bacteroides conjugal elements. J Bacteriol. 1988 Mar;170(3):1319–1324. doi: 10.1128/jb.170.3.1319-1324.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Yanisch-Perron C., Vieira J., Messing J. Improved M13 phage cloning vectors and host strains: nucleotide sequences of the M13mp18 and pUC19 vectors. Gene. 1985;33(1):103–119. doi: 10.1016/0378-1119(85)90120-9. [DOI] [PubMed] [Google Scholar]