Recognition of GT mismatches by Vsr mismatch endonuclease

Fox, Keith R. and Allinson, Sarah L. and Sahagun-Krause, Heidi and Brown, Tom (2000) Recognition of GT mismatches by Vsr mismatch endonuclease. Nucleic Acids Research, 28 (13). pp. 2535-2540. ISSN 0305-1048

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Abstract

The Vsr mismatch endonuclease recognises the sequence CTWGG (W = A or T) in which the underlined thymine is paired with guanine and nicks the DNA backbone on the 5'-side of the mispaired thymine. By using base analogues of G and T we have explored the functional groups on the mismatch pair which are recognised by the enzyme. Removal of the thymine 5-methyl group causes a 60% reduction in activity, while removing the 2-amino group of guanine reduces cleavage by 90%. Placing 5-aminopurine or nebularine opposite T generates mismatches which are cut at a much lower rate (0.1%). When either base is removed, generating a pseudoabasic site (1',2'-dideoxyribose), the enzyme still produces site-specific cleavage, but at only 1% of the original rate. Although TT and CT mismatches at this position are cleaved at a low rate (~ 1%), mismatches with other bases (such as GA and AC) and Watson-Crick base pairs are not cleaved by the enzyme. There is also no cleavage when the mismatched T is replaced with difluorotoluene.

Item Type:
Journal Article
Journal or Publication Title:
Nucleic Acids Research
Uncontrolled Keywords:
/dk/atira/pure/subjectarea/asjc/1300/1311
Subjects:
ID Code:
154528
Deposited By:
Deposited On:
05 May 2021 14:25
Refereed?:
Yes
Published?:
Published
Last Modified:
09 Jun 2021 05:56