Cleavage of a DNA–RNA–DNA/DNA chimeric substrate containing a single ribonucleotide at the DNA–RNA junction with prokaryotic RNases HII
Open Access
- 29 October 2002
- journal article
- research article
- Published by Wiley in FEBS Letters
- Vol. 531 (2) , 204-208
- https://doi.org/10.1016/s0014-5793(02)03503-2
Abstract
We have analyzed the cleavage specificities of various prokaryotic Type 2 ribonucleases H (RNases H) on chimeric DNA–RNA–DNA/DNA substrates containing one to four ribonucleotides. RNases HII from Bacillus subtilis and Thermococcus kodakaraensis cleaved all of these substrates to produce a DNA segment with a 5′‐monoribonucleotide. Consequently, these enzymes cleaved even the chimeric substrate containing a single ribonucleotide at the DNA–RNA junction (5′‐side of the single ribonucleotide). In contrast, Escherichia coli RNase HI and B. subtilis RNase HIII did not cleave the chimeric substrate containing a single ribonucleotide. These results suggest that bacterial and archaeal RNases HII are involved in excision of a single ribonucleotide misincorporated into DNA.Keywords
This publication has 26 references indexed in Scilit:
- Importance of an N-terminal extension in ribonuclease HII from Bacillus stearothermophilus for substrate bindingJournal of Bioscience and Bioengineering, 2002
- Archaeoglobus fulgidus RNase HII in DNA Replication: Enzymological Functions and Activity Regulation via Metal CofactorsBiochemical and Biophysical Research Communications, 2001
- Catalytic center of an archaeal type 2 ribonuclease H as revealed by X‐ray crystallographic and mutational analysesProtein Science, 2001
- Structural biochemistry of a type 2 RNase H: RNA primer recognition and removal during DNA replication11Edited by K. MorikawaJournal of Molecular Biology, 2001
- Identification of the Genes Encoding Mn2+-Dependent RNase HII and Mg2+-Dependent RNase HIII from Bacillus subtilis: Classification of RNases H into Three FamiliesBiochemistry, 1998
- Kinetic Analysis of Escherichia Coli Ribonuclease HI Using Oligomeric DNA/RNA Substrates Suggests an Alternative Mechanism for the Interaction between the Enzyme and the SubstrateEuropean Journal of Biochemistry, 1995
- Functions and Structures of Ribonuclease H EnzymesPublished by Springer Nature ,1995
- Structure of Ribonuclease H Phased at 2 Å Resolution by MAD Analysis of the Selenomethionyl ProteinScience, 1990
- Three-dimensional structure of ribonuclease H from E. coliNature, 1990
- Function of RNase H in DNA replication revealed by RNase H defective mutants of Escherichia coliMolecular Genetics and Genomics, 1984