ATPase Activity of the Type IC Restriction-Modification SystemEcoR124II |
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| Institution: | 1. USDA-ARS Edward T. Schafer Agricultural Research Center, Fargo, ND, USA;2. South Dakota State University, Brookings, SD, USA;3. University of Guelph, Ontario, Canada;1. School of Applied Biosciences, Kyungpook National University, Daegu 702-701, Republic of Korea;2. International Institute of Agricultural Research & Development, Kyungpook National University, Daegu 702-701, Republic of Korea |
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| Abstract: | We have investigated the ATPase activity of the type IC restriction-modification (R – M) systemEcoR124II. As with all type I R – M systemsEcoR 124II requires ATP hydrolysis to cut DNA. We determined theKMfor ATP to be 10?5to 10?4M. By measuring ATP hydrolysis under different conditions and by simultaneously monitoring DNA restriction, methylation and ATP hydrolysis we propose that the order of events during restriction is: (1) binding ofEcoR124II to a non-methylated recognition sequence, (2) start of DNA-dependent ATP hydrolysis which continues even after restriction is complete, (3) restriction of DNA, (4) methylation of the product. Non-cleavable DNA substrates, such as recognition site containing oligonucleotides, also support ATP hydrolysis. Methylation can also occur prior to ATP hydrolysis and prevent DNA degradation. |
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