Raghavendra, Nidhanapati K. ; Rao, Desirazu N. (2005) Exogenous AdoMet and its analogue sinefungin differentially influence DNA cleavage by R.EcoP15I-Usefulness in SAGE Biochemical and Biophysical Research Communications, 334 (3). pp. 803-811. ISSN 0006-291X
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Official URL: http://linkinghub.elsevier.com/retrieve/pii/S0006-...
Related URL: http://dx.doi.org/10.1016/j.bbrc.2005.06.171
Abstract
While it has been demonstrated that AdoMet is required for DNA cleavage by Type III restriction enzymes, here we show that in the presence of exogenous AdoMet, the head-to-head oriented recognition sites are cleaved only on a supercoiled DNA. On a linear DNA, exogenous AdoMet strongly drives methylation while inhibiting cleavage reaction. Strikingly, AdoMet analogue sinefungin results in cleavage at all recognition sites irrespective of the topology of DNA. The cleavage reaction in the presence of sinefungin is ATP dependent. The site of cleavage is comparable with that in the presence of AdoMet. The use of EcoP15I restriction in presence of sinefungin as an improved tool for serial analysis of gene expression is discussed.
| Item Type: | Article |
|---|---|
| Source: | Copyright of this article belongs to Elsevier Science. |
| Keywords: | AdoMet; Sinefungin; EcoP15I; DNA Topology; Site Requirement; SAGE |
| ID Code: | 51219 |
| Deposited On: | 28 Jul 2011 07:26 |
| Last Modified: | 28 Jul 2011 07:26 |
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