P1 nuclease cleavage is dependent on length of the mismatches in DNA.
P1 nuclease is one of the most extensively used single-strand DNA specific nucleases in molecular biology. In modern biology, it is used as an enzymatic probe to detect altered DNA conformations. It is well documented that P1 cleaves single-stranded nucleic acids and single-stranded DNA regions. The fact that P1 can act under a wide range of conditions, including physiological pH and temperature make it the most commonly used enzymatic probe in DNA structural studies. Surprisingly, to this date, there is no study to characterize the influence of length of mismatches on P1 sensitivity. Using a series of radioactively labeled oligomeric DNA substrates-containing mismatches, we find that P1 nuclease cleavage is dependent on the length of mismatches. P1 does not cleave DNA when there is a single-base mismatch. P1 cleavage efficiency is optimum when mismatch length is 3 nt or more. Changing the position of the mismatches also does not make any difference in cleavage efficacy. These novel findings on P1 properties have implications for its use in DNA structure and DNA repair studies.
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Related Subject Headings
- Potassium Permanganate
- Hydrolysis
- Genomic Instability
- Developmental Biology
- Deoxyribonucleases
- DNA Primers
- Base Sequence
- Base Pair Mismatch
- 3101 Biochemistry and cell biology
- 0601 Biochemistry and Cell Biology
Citation
Published In
DOI
EISSN
ISSN
Publication Date
Volume
Issue
Start / End Page
Related Subject Headings
- Potassium Permanganate
- Hydrolysis
- Genomic Instability
- Developmental Biology
- Deoxyribonucleases
- DNA Primers
- Base Sequence
- Base Pair Mismatch
- 3101 Biochemistry and cell biology
- 0601 Biochemistry and Cell Biology