Comparative study of class 1 integron and Vibrio cholerae superintegron integrase activities.
Superintegrons (SIs) and multiresistant integrons (MRIs) have two main structural differences: (i) the SI platform is sedentary, while the MRI platform is commonly associated with mobile DNA elements and (ii) the recombination sites (attC) of SI gene cassette clusters are highly homogeneous, while those of MRI cassette arrays are highly variable in length and sequence. In order to determine if the latter difference was correlated with a dissimilarity in the recombination activities, we conducted a comparative study of the integron integrases of the class 1 MRI (IntI1) and the Vibrio cholerae SI (VchIntIA). We developed two assays that allowed us to independently measure the frequencies of cassette deletion and integration at the cognate attI sites. We demonstrated that the range of attC sites efficiently recombined by VchIntIA is narrower than the range of attC sites efficiently recombined by IntI1. Introduction of mutations into the V. cholerae repeats (VCRs), the attC sites of the V. cholerae SI cassettes, allowed us to map positions that affected the VchIntIA and IntI1 activities to different extents. Using a cointegration assay, we established that in E. coli, attI1-x-VCR recombination catalyzed by IntI1 was 2,600-fold more efficient than attIVch-x-VCR recombination catalyzed by VchIntIA. We performed the same experiments in V. cholerae and established that the attIVch-x-VCR recombination catalyzed by VchIntIA was 2,000-fold greater than the recombination measured in E. coli. Taken together, our results indicate that in the V. cholerae SI, the substrate recognition and recombination reactions mediated by VchIntIA might differ from the class 1 MRI paradigm.
Biskri
Latefa
L
Unité Postulante Plasticité du Génome Bactérien, CNRS URA 2171, Département Structure et Dynamique des Génomes, Institut Pasteur, 25 rue du Dr Roux, 75724, Paris, France.
Bouvier
Marie
M
Guérout
Anne-Marie
AM
Boisnard
Stéphanie
S
Mazel
Didier
D
D001483:Base Sequence; D024901:Drug Resistance, Multiple, Bacterial / Q000235:genetics; D004926:Escherichia coli / Q000235:genetics; D015964:Gene Expression Regulation, Bacterial; D019426:Integrases / Q000378:metabolism*; D032023:Integrons / Q000502:physiology*; D008969:Molecular Sequence Data; D016254:Mutagenesis, Insertional; D011995:Recombination, Genetic; D016415:Sequence Alignment; D012689:Sequence Homology, Nucleic Acid; D014734:Vibrio cholerae / Q000201:enzymology* / Q000235:genetics*
D019426:Integrases; C115006:integron integrase IntI1
Proc Natl Acad Sci U S A. 2001 Jan 16;98(2):652-7
11209061
J Bacteriol. 2003 Mar;185(6):2036-41
12618471
Curr Opin Microbiol. 2001 Oct;4(5):565-9
11587934
Appl Environ Microbiol. 2001 Nov;67(11):5240-6
11679351
Environ Microbiol. 2003 May;5(5):383-94
12713464
Appl Environ Microbiol. 2004 Feb;70(2):1160-8
14766601
Clin Microbiol Infect. 2004 Apr;10(4):272-88
15059115
Mol Gen Genet. 1988 Feb;211(2):320-5
2832705
Annu Rev Biochem. 1989;58:913-49
2528323
EMBO J. 1990 Apr;9(4):1275-81
2157593
Gene. 1991 Jun 15;102(1):75-8
1840539
J Bacteriol. 1991 Sep;173(17):5523-31
1885529
J Bacteriol. 1992 Mar;174(5):1574-85
1311297
Genetica. 1993;90(2-3):115-32
8119588
Gene. 1994 Jan 28;138(1-2):1-7
8125283
Gene. 1994 May 3;142(1):49-54
8181756
Mol Microbiol. 1994 Apr;12(2):241-51
8057849
J Bacteriol. 1994 Sep;176(17):5450-8
8071223
J Bacteriol. 1995 Jul;177(14):4121-30
7608087
Microbiology. 1995 Dec;141 ( Pt 12):3015-27
8574395
EMBO J. 1996 Mar 15;15(6):1203-10
8635452
Ciba Found Symp. 1997;207:192-202; discussion 202-5
9189642
Mol Microbiol. 1997 Nov;26(3):441-53
9402016
Nucleic Acids Res. 1998 Jan 15;26(2):391-406
9421491
Mol Microbiol. 1997 Nov;26(4):731-45
9427403
Science. 1998 Apr 24;280(5363):605-8
9554855
Mol Microbiol. 1998 Jul;29(2):477-90
9720866
APMIS Suppl. 1998;84:37-42
9850680
Ann N Y Acad Sci. 1999 May 18;870:68-80
10415474
J Biol Chem. 1999 Nov 12;274(46):33105-13
10551881
Infect Immun. 2000 Nov;68(11):6391-7
11035750
FEMS Microbiol Lett. 2001 Feb 5;195(1):59-65
11166996
Mol Microbiol. 2001 Nov;42(3):587-601
11722728
J Bacteriol. 2002 Mar;184(6):1712-21
11872723
J Bacteriol. 2002 Mar;184(6):1811-5
11872738
Mol Microbiol. 2002 Mar;43(6):1657-69
11952913
J Bacteriol. 2002 Jun;184(11):3017-26
12003943
Int J Med Microbiol. 2002 Jul;292(2):115-25
12195734
J Bacteriol. 2003 Feb;185(3):918-28
12533467
Genome Res. 2003 Mar;13(3):428-42
12618374
J Bacteriol. 2001 Apr;183(8):2535-42
11274113