Arm site independence of coliphage HK022 integrase in human cells

被引:0
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作者
Natalia Malchin
Chen N. Tuby
Ezra Yagil
Mikhail Kolot
机构
[1] Tel-Aviv University,Department of Biochemistry and Molecular Biology
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关键词
Coliphage HK022; Integrase; Site-specific recombination; Human embryonic kidney cell culture;
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摘要
The integrase encoded by the lambdoid phage HK022 (Int-HK022) resembles its coliphage λ counterpart (Int-λ) in the roles of the cognate DNA arm binding sites and in controlling the direction of the reaction. We show here that within mammalian cells, Int-HK022 does not exhibit such a control. Rather, Int-HK022 recombined between all ten possible pairwise att site combinations, including attB × attB that was more effective than the conventional integrative attP × attB reaction. We further show that Int-HK022 depends on the accessory integration host factor (IHF) protein considerably less than Int-λ and exhibits stronger binding affinity to the att core. These differences explain why wild-type Int-HK022 is active in mammalian cells whereas Int-λ is active there only as an IHF-independent mutant.
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页码:403 / 413
页数:10
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