PMID:9346880
| Citation |
Lima, WF and Crooke, ST (1997) Cleavage of single strand RNA adjacent to RNA-DNA duplex regions by Escherichia coli RNase H1. J. Biol. Chem. 272:27513-6 |
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| Abstract |
RNase H1 from Escherichia coli cleaves single strand RNA extending 3' from an RNA-DNA duplex. Substrates consisting of a 25-mer RNA annealed to complementary DNA ranging in length from 9-17 nucleotides were designed to create overhanging single strand RNA regions extending 5' and 3' from the RNA-DNA duplex. Digestion of single strand RNA was observed exclusively within the 3' overhang region and not the 5' overhang region. RNase H digestion of the 3' overhang region resulted in digestion products with 5'-phosphate and 3'-hydroxyl termini. The number of single strand RNA residues cleaved by RNase H is influenced by the sequence of the single strand RNA immediately adjacent to the RNA-DNA duplex and appears to be a function of the stacking properties of the RNA residues adjacent to the RNA-DNA duplex. RNase H digestion of the 3' overhang region was not observed for a substrate that contained a 2'-methoxy antisense strand. The introduction of 3 deoxynucleotides at the 5' terminus of the 2'-methoxy antisense oligonucleotide resulted in cleavage. These results offer additional insights into the binding directionality of RNase H with respect to the heteroduplex substrate. |
| Links | |
| Keywords |
DNA/chemistry; Escherichia coli/enzymology; Hydrolysis; Nucleic Acid Heteroduplexes; RNA/chemistry; RNA/metabolism; Ribonuclease H/metabolism |
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