Journal
JOURNAL OF BIOCHEMISTRY
Volume 154, Issue 6, Pages 505-511Publisher
OXFORD UNIV PRESS
DOI: 10.1093/jb/mvt080
Keywords
DNA sequence or structure; endonuclease activity; mismatch repair; MutL; Pseudomonas aeruginosa
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Funding
- Secretaria de Ciencia y Tecnologia, Universidad Nacional de Cordoba
- Ministerio de Ciencia, Tecnologia e Innovacion de la Provincia de Cordoba
- Agencia Nacional de Promocion Cientifica y Tecnologica
- Consejo Nacional de Investigaciones Cientificas y Tecnicas
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The hallmark of the mismatch repair system in bacterial and eukaryotic organisms devoid of MutH is the presence of a MutL homologue with endonuclease activity. The aim of this study was to analyse whether different DNA structures affect Pseudomonas aeruginosa MutL (PaMutL) endonuclease activity and to determine if a specific nucleotide sequence is required for this activity. Our results showed that PaMutL was able to nick covalently closed circular plasmids but not linear DNA at high ionic strengths, while the activity on linear DNA was only found below 60mM salt. In addition, single strand DNA, ss/ds DNA boundaries and negatively supercoiling degree were not required for PaMutL nicking activity. Finally, the analysis of the incision sites revealed that PaMutL, as well as Bacillus thuringiensis MutL homologue, did not show DNA sequence specificity.
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