Recognition of GT mismatches by Vsr mismatch endonuclease
Recognition of GT mismatches by Vsr mismatch endonuclease
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 2-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.
escherichia-coli k-12, short patch repair, cytosine methylation, crystal-structure, dna duplex, gene, 5-methyl-cytosine, difluorotoluene, cleavage, adenine
2535-2540
Fox, K.R.
9da5debc-4e45-473e-ab8c-550d1104659f
Allinson, S.L.
dc35efa2-6bfe-41b6-a89f-812f7070e151
Sahagun-Krause, H.
d7ca266c-61f0-4864-8b86-6aa2e731275b
Brown, T.
a64aae36-bb30-42df-88a2-11be394e8c89
1 July 2000
Fox, K.R.
9da5debc-4e45-473e-ab8c-550d1104659f
Allinson, S.L.
dc35efa2-6bfe-41b6-a89f-812f7070e151
Sahagun-Krause, H.
d7ca266c-61f0-4864-8b86-6aa2e731275b
Brown, T.
a64aae36-bb30-42df-88a2-11be394e8c89
Fox, K.R., Allinson, S.L., Sahagun-Krause, H. and Brown, T.
(2000)
Recognition of GT mismatches by Vsr mismatch endonuclease.
Nucleic Acids Research, 28 (13), .
(doi:10.1093/nar/28.13.2535).
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 2-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.
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More information
Published date: 1 July 2000
Keywords:
escherichia-coli k-12, short patch repair, cytosine methylation, crystal-structure, dna duplex, gene, 5-methyl-cytosine, difluorotoluene, cleavage, adenine
Identifiers
Local EPrints ID: 18926
URI: http://eprints.soton.ac.uk/id/eprint/18926
ISSN: 0305-1048
PURE UUID: b968ee00-4197-4730-94e5-7b28c815f88b
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Date deposited: 19 Jan 2006
Last modified: 16 Mar 2024 02:36
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Author:
S.L. Allinson
Author:
H. Sahagun-Krause
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