Ribonuclease H (RNase H) plays crucial roles in replication of retroviruses, including the human immunodeficiency virus (HIV) as well as involvement in normal cellular events including DNA replication. The purpose of this project is to gain a detailed understanding of the precise nature of the substrates for RNases H, the chemical makeup of cellular RNases H. The enzymes derived from cellular sources have higher specific activities than those of retroviral origin and have somewhat different specificities. We have found that separation of the RNase H domain of MuLV reverse transcriptase from the polymerase domain creates an active enzyme with specificities different from the parental RNase H but still with low specific activity. If we produce RNases H from yeasts, they have high specific activities regardless of their association with their own special domains. These results indicate that the differences in these proteins reside in the RNase H domain itself.
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| Budihas, Scott R; Gorshkova, Inna; Gaidamakov, Sergei et al. (2005) Selective inhibition of HIV-1 reverse transcriptase-associated ribonuclease H activity by hydroxylated tropolones. Nucleic Acids Res 33:1249-56 |
| Gaidamakov, Sergei A; Gorshkova, Inna I; Schuck, Peter et al. (2005) Eukaryotic RNases H1 act processively by interactions through the duplex RNA-binding domain. Nucleic Acids Res 33:2166-75 |
| Nowotny, Marcin; Gaidamakov, Sergei A; Crouch, Robert J et al. (2005) Crystal structures of RNase H bound to an RNA/DNA hybrid: substrate specificity and metal-dependent catalysis. Cell 121:1005-16 |
| Chan, King C; Budihas, Scott R; Le Grice, Stuart F J et al. (2004) A capillary electrophoretic assay for ribonuclease H activity. Anal Biochem 331:296-302 |
| Jeong, Ho-Sang; Backlund, Peter S; Chen, Hao-Chia et al. (2004) RNase H2 of Saccharomyces cerevisiae is a complex of three proteins. Nucleic Acids Res 32:407-14 |
| Broccoli, Sonia; Rallu, Fabien; Sanscartier, Patrick et al. (2004) Effects of RNA polymerase modifications on transcription-induced negative supercoiling and associated R-loop formation. Mol Microbiol 52:1769-79 |
| Backer, Marina V; Gaynutdinov, Timur I; Gorshkova, Inna I et al. (2003) Humanized docking system for assembly of targeting drug delivery complexes. J Control Release 89:499-511 |
| Pileur, Frederic; Andreola, Marie-Line; Dausse, Eric et al. (2003) Selective inhibitory DNA aptamers of the human RNase H1. Nucleic Acids Res 31:5776-88 |
| Cerritelli, Susana M; Frolova, Ella G; Feng, Chiguang et al. (2003) Failure to produce mitochondrial DNA results in embryonic lethality in Rnaseh1 null mice. Mol Cell 11:807-15 |
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