Open AccessInhibition of IS 2 transposition by factor for inversion stimulation
Author(s) -
Lei GuangSheng,
Chen ChiiJaan,
Yuan Hanna S.,
Wang ShaoHung,
Hu ShiauTing
Publication year - 2007
Publication title -
fems microbiology letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.899
H-Index - 151
eISSN - 1574-6968
pISSN - 0378-1097
DOI - 10.1111/j.1574-6968.2007.00864.x
Subject(s) - microbiology and biotechnology , promoter , transposase , dna footprinting , transposable element , transcription (linguistics) , binding site , biology , transcription factor , mutant , gene , dna binding protein , genetics , gene expression , linguistics , philosophy
The effect of factor for inversion stimulation (Fis) protein on IS 2 transposition was investigated. A full‐length IS 2 was found to transpose at a frequency 64 times lower in a normal Escherichia coli than in a fis − mutant. To investigate whether Fis affects IS 2 transposition by DNA binding, gel retardation and DNase I footprinting experiments were performed. Analysis of Fis binding to the left terminus of IS 2 revealed that Fis binds to nucleotide number 44–60 located between the −35 and −10 regions of the major IS 2 promoter. To further determine whether Fis binding affects IS 2 transcription, the major IS 2 promoter was fused to a luciferase gene and assayed for its transcription efficiency in the presence or absence of Fis. The results showed that Fis reduced transcription from the major IS 2 promoter by approximately sixfold. Analysis of Fis binding to the right terminal repeat of IS 2 revealed that Fis binds to the inner end of the repeat, which is the same region as the place where the IS 2 transposase binds. These results suggest that Fis inhibits IS 2 transposition by blocking the binding sites of IS 2 transposase and by repressing the transcription of IS 2 genes.