DNA complexes obtained with the integron integrase IntI1 at the attI1 site

被引:49
作者
Gravel, A
Fournier, B
Roy, PH
机构
[1] CHU Laval, Ctr Rech Infectiol, Quebec City, PQ G1V 4G2, Canada
[2] Univ Laval, Fac Sci & Genie, Dept Biochim, Quebec City, PQ G1V 4G2, Canada
基金
英国医学研究理事会;
关键词
D O I
10.1093/nar/26.19.4347
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
Integrons are genetic elements that are able to capture genes by a site-specific recombination mechanism. Integrons contain a gene coding for a lambda-like integrase that carries out site-specific recombination by interacting with two different target sites; the attI site and the palindromic sequence attC (59 base element). Cassette integrations usually involve the attI site, while cassette excisions use attC. Therefore, the integrase should bind both sites to cleave DNA and perform site-specific recombination reactions. We have used purified maltose-binding protein fused with the integrase (MBP-IntI1) and native IntI1 protein and gel retardation assays with fragments containing the complete and partial attI1 site to show formation of four complexes in this region. Chemical modification of specific nucleotides within the attI1 site was used to investigate their interference with binding of the integrase protein. We attribute IntI1 specific binding to four regions in the attI1 site and a GTTA consensus sequence is found in three of the four regions. Interference by modified guanine and thymine residues in the DNA major groove and adenine residues in the minor groove were observed, indicating that the integrase interacts with both sides of the helix. Binding of IntI1 to attC is also discussed.
引用
收藏
页码:4347 / 4355
页数:9
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