Detection of an IS2-encoded 46-kilodalton Protein Capable of Binding Terminal Repeats of IS2

Overview

Journal J Bacteriol

Publisher American Society for Microbiology

Specialty Microbiology

Date 1996 Oct 1

PMID 8824609

Citations 8

Authors

S T Hu

L C Lee

G S Lei

Affiliations

Soon will be listed here.

Abstract

The genome of the transposable element IS2 contains five open reading frames that are capable of encoding proteins greater than 50 amino acids; however, only one IS2 protein of 14 kDa had been detected. By replacing the major IS2 promoter located in the right terminal repeat of IS2 with the T7 promoter to express IS2 genes, we have detected another IS2 protein of 46 kDa. This 46-kDa protein was designated InsAB'. Analyses of the InsAB' sequence revealed motifs that are characteristic of transposases of other transposable elements. InsAB' has the ability to bind both terminal repeat sequences of IS2. It was shown to bind a 27-bp sequence (5'-GTTAAGTGATAACAGATGTCTGGAAAT-3', positions 1316 to 1290 by our numbering system [16 to 42 by the previous numbering system]) located at the inner end of the right terminal repeat and a 31-bp sequence (5'-TTATTTAAGTGATATTGGTTGTCTGGAGATT-3', positions 46 to 16 [1286 to 1316]), including the last 27 bp of the inner end and the adjacent 4 bp of the left terminal repeat of IS2. This result suggests that InsAB' is a transposase of IS2. Since there is no open reading frame capable of encoding a 46-kDa protein in the entire IS2 genome, this 46-kDa protein is probably produced by a translational frameshifting mechanism.

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References

Prere M, Chandler M, Fayet O . Transposition in Shigella dysenteriae: isolation and analysis of IS911, a new member of the IS3 group of insertion sequences. J Bacteriol. 1990; 172(7):4090-9. PMC: 213396. DOI: 10.1128/jb.172.7.4090-4099.1990. View

Sekine Y, Ohtsubo E . DNA sequences required for translational frameshifting in production of the transposase encoded by IS1. Mol Gen Genet. 1992; 235(2-3):325-32. PMC: 7088269. DOI: 10.1007/BF00279377. View

Laemmli U . Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970; 227(5259):680-5. DOI: 10.1038/227680a0. View

Jacks T, Madhani H, Masiarz F, Varmus H . Signals for ribosomal frameshifting in the Rous sarcoma virus gag-pol region. Cell. 1988; 55(3):447-58. PMC: 7133365. DOI: 10.1016/0092-8674(88)90031-1. View

Ghosal D, Sommer H, Saedler H . Nucleotide sequence of the transposable DNA-element IS2. Nucleic Acids Res. 1979; 6(3):1111-22. PMC: 327757. DOI: 10.1093/nar/6.3.1111. View

Amemura J, Ichikawa H, Ohtsubo E . Tn3 transposition immunity is conferred by the transposase-binding domain in the terminal inverted-repeat sequence of Tn3. Gene. 1990; 88(1):21-4. DOI: 10.1016/0378-1119(90)90055-v. View

Fiandt M, SZYBALSKI W, MALAMY M . Polar mutations in lac, gal and phage lambda consist of a few IS-DNA sequences inserted with either orientation. Mol Gen Genet. 1972; 119(3):223-31. DOI: 10.1007/BF00333860. View

Joos H, Timmerman B, Montagu M, Schell J . Genetic analysis of transfer and stabilization of Agrobacterium DNA in plant cells. EMBO J. 1983; 2(12):2151-60. PMC: 555427. DOI: 10.1002/j.1460-2075.1983.tb01716.x. View

Polard P, Prere M, Chandler M, Fayet O . Programmed translational frameshifting and initiation at an AUU codon in gene expression of bacterial insertion sequence IS911. J Mol Biol. 1991; 222(3):465-77. DOI: 10.1016/0022-2836(91)90490-w. View

10.

Rezsohazy R, Hallet B, Mahillon J, Delcour J . IS231V and W from Bacillus thuringiensis subsp. israelensis, two distant members of the IS231 family of insertion sequences. Plasmid. 1993; 30(2):141-9. DOI: 10.1006/plas.1993.1041. View

11.

Weiss R, Dunn D, Shuh M, Atkins J, GESTELAND R . E. coli ribosomes re-phase on retroviral frameshift signals at rates ranging from 2 to 50 percent. New Biol. 1989; 1(2):159-69. View

12.

Johnson R, Reznikoff W . DNA sequences at the ends of transposon Tn5 required for transposition. Nature. 1983; 304(5923):280-2. DOI: 10.1038/304280a0. View

13.

Zerbib D, Prentki P, Gamas P, Freund E, Galas D, Chandler M . Functional organization of the ends of IS1: specific binding site for an IS 1-encoded protein. Mol Microbiol. 1990; 4(9):1477-86. View

14.

Rezsohazy R, Hallet B, Delcour J, Mahillon J . The IS4 family of insertion sequences: evidence for a conserved transposase motif. Mol Microbiol. 1993; 9(6):1283-95. DOI: 10.1111/j.1365-2958.1993.tb01258.x. View

15.

Sekine Y, Nagasawa H, Ohtsubo E . Identification of the site of translational frameshifting required for production of the transposase encoded by insertion sequence IS 1. Mol Gen Genet. 1992; 235(2-3):317-24. PMC: 7088211. DOI: 10.1007/BF00279376. View

16.

Zerbib D, Polard P, Escoubas J, Galas D, Chandler M . The regulatory role of the IS1-encoded InsA protein in transposition. Mol Microbiol. 1990; 4(3):471-7. DOI: 10.1111/j.1365-2958.1990.tb00613.x. View

17.

Wiegand T, Reznikoff W . Interaction of Tn5 transposase with the transposon termini. J Mol Biol. 1994; 235(2):486-95. DOI: 10.1006/jmbi.1994.1008. View

18.

Garner M, Revzin A . A gel electrophoresis method for quantifying the binding of proteins to specific DNA regions: application to components of the Escherichia coli lactose operon regulatory system. Nucleic Acids Res. 1981; 9(13):3047-60. PMC: 327330. DOI: 10.1093/nar/9.13.3047. View

19.

Derbyshire K, Hwang L, Grindley N . Genetic analysis of the interaction of the insertion sequence IS903 transposase with its terminal inverted repeats. Proc Natl Acad Sci U S A. 1987; 84(22):8049-53. PMC: 299474. DOI: 10.1073/pnas.84.22.8049. View

20.

Chandler M, Fayet O . Translational frameshifting in the control of transposition in bacteria. Mol Microbiol. 1993; 7(4):497-503. DOI: 10.1111/j.1365-2958.1993.tb01140.x. View