The retroviral RNase H was discovered by Karin Moelling during her thesis work at the Max-Planck-Institute of Virology in Tuebingen, Germany. She was trying to isolate the enzyme required for replication of the avian myoloblastosis virus, a chicken retrovirus. She looked for an RNA-dependent RNA polymerase and when she heard from Howard Temin’s work on the discovery of a reverse transcriptase, she just had to exchange four NTPs to four dNTPs to obtain millions of counts!! Then she discoverd the RNase H as an essential function for retroviral replication. Later on the enzyme was also found in HIV and characterized in detail in respect to molecular mechanisms and target validation. Based on this a therapeutic approach against HIV was developed by use of a hairpin loop-structured oligodeoxynucleotide, which was published in Nature Biotechnology in 2007 and described as driving HIV into suicide. The mechanism is based on the activation of the RNase H by an oligonucleotide, which destroys the viral RNA and infectivity in cell-free particles. It would be useful to prevent sexual transmission in form of a microbicide or mother-to-child transmission. The method can be described as RNA silencing by siDNA as a follow-up of siRNA and can be applied to various viruses and cancer.
Most important recent papers for immediate reading
- Matzen, K., Elzaouk, L., Matskevich A.A., Nitzsche, A., Heinrich, J. and Moelling, K. RNase H-mediated retrovirus destruction in vivo triggered by oligodeoxynucleotides. Nature Biotechnol. 25, 669-674 (2007). Matzen 2007.pdf
- Editorial Comment: Assisted suicide for retroviruses Nature Biotechnol. 25, 643-644(2007) – Comment Matzen 2007.pdf
Publications about RNase H with selected pdf files
- Moelling, K., Bolognesi, D.P., Bauer, H., Büsen, W., Plassmann, H.W. and Hausen, P. Association of the viral reverse transcriptase with an enzyme degrading the RNA moiety of RNA-DNA hybrids. Nature New Biology 234, 240-243 (1971) Comparative Leukemia Research, Bibl. Haematol. 39, 536-550 (1973).
- Moelling, K.: Characterization of Reverse Transcriptase and RNase H from Friend murine leukemia virus. Virology 62, 46-59 (1974b).
- Moelling, K. Reverse transcriptase and RNase H: Present in a murine virus and in both subunits of an avian virus. Cold Spring Harbor Symp. Quant. Biol. 39, 969-973 (1974a).
- Moelling, K. Exciting days over for reverse transcriptase? Nature 256, 258-260 (1975).
- Moelling, K. Comparison between an avian and a murine viral reverse transcriptase RNase H complex. Comparative Leukemia Res., Bibl. Haematol. 43, 121-124 (1976).
- Moelling, K. Further characterization of the Friend murine leukemia virus RNase H complex. J. Virol. 18, 418-425 (1976).
- Moelling, K. Anything new about reverse transcriptase? Nature 264, 111-112 (1976)
- Friedrich, R. and Moelling, K. Effect of viral RNase H on the avian sarcoma viral genome during early transcription in vitro. J. Virol. 31, 630-638 (1979).
- Moelling, K. and Friis, R.R. Two avian sarcoma virus mutants with defects in the DNA polymerase-RNase H complex. J. Virol. 32, 370-378 (1979).
- Sykora, K.W. and Moelling, K. Properties of the avian viral protein p12. J. Gen. Virol. 55, 379-391 (1981).
- Hansen, J., Schulze, T. and Moelling, K. RNase H activity associated with bacterially expressed reverse transcriptase of human T-cell lymphotropic virus III/lymphadenopathy-associated virus. J. Biol. Chem. 262, 12393-12396 (1987).
- Hansen, J., Schulze, T., Mellert, W. and Moelling, K. Identification and characterization of HIV-specific RNase H by monoclonal antibody. EMBO J. 7, 239-243 (1988). – Hansen 1988.pdf
- Moelling, K., Schulze, T. and Diringer, H. Inhibition of HIV-1 RNase H by sulfated polyanions. J. Virol. 63, 5489-5492 (1989).
- Wöhrl, B. and Moelling, K. Interaction of HIV-1 RNase H with polypurine tract containing RNA-DNA hybrids. Biochem. 29, 10141-10147 (1990). – Wöhrl 1990.pdf
- Wöhrl, B.M., Moelling, K. Coupling of reverse transcriptase and RNase H during HIV-1 replication. Behring Inst Mitt. (1991)
- Tisdale, M., Schulze, T., Larder, B.A. and Moelling, K. Mutations within the RNase H domain of HIV-1 reverse transcriptase abolish virus infectivity. J. Gen. Virol. 72, 59-66 (1991) – Tisdale 1991
- Schulze, T., Nawrath, M. and Moelling, K. Cleavage of the HIV-1 p66 reverse transcriptase/RNase H by the p9 protease in vitro generates active p15 RNase H. Archives of Virol. 118, 179-188 (1991).
- Wöhrl, B., Volkmann, S. and Moelling, K. Mutations of a conserved residue within HIV-1 ribonuclease H affect its exo- and endonuclease activities. J. Mol. Biol. 220, 801-818 (1991).
- Volkmann, S., Dannull, J., Moelling, K. The polypurine tract, PPT, of HIV as target for antisense and triple-helix-forming oligonucleotides. Biochimie 75, 71-78 (1993).
- Volkmann, S., Wöhrl, B.M., Tisdale, M. and Moelling, K. Enzymatic analysis of two HIV-1 reverse transcriptase mutants with mutations in carboxyl-terminal amino acid residues conserved among retroviral ribonucleases H. J. Biol. Chem. 268, 2674-2683 (1993).
- Volkmann, S., Jendis, J., Frauendorf, A. and Moelling, K. Inhibition of HIV-1 reverse transcription by triple-helix forming oligonucleotides with viral RNA. Nucl. Acid. Res. 23, 1204-1212 (1995) – Volkmann 1995
- Jendis, J., Strack, B., Volkmann, S., Böni, J. and Moelling, K. (1996) Inhibition of Replication of Fresh HIV Type 1 Patient Isolates by a Polypurine Tract-Specific Self-Complementary Oligodeoxynucleotide. AIDS Res Hum Retroviruses 12 (12):1161-8 – Jendis 1996
- Jendis, J., Strack, B. and Moelling, K. (1998) Inhibition of replication of drug-resistant HIV type 1 isolates by polypurine tract-specific oligodeoxynucleotide TFO A. AIDS Res Hum Retroviruses. 14 (11): 999-1005 – Jendis 1998
- Moelling, K., Abels, S., Jendis, J., Matskevich, A., Heinrich, J. (2006) Silencing of HIV by hairpin-loop-structured DNA oligonucleotide. FEBS Letters 580: 3545–3550 – Moelling 2006
- Matskevich, A.A., Ziogas, A., Heinrich, J., Quast, S.A. and Moelling, K. Short Partially Double-Stranded Oligodeoxynucleotide Induces Reverse Transcriptase/RNase H-Mediated Cleavage of HIV RNA and Contributes to Abrogation of Infectivity of Virions. AIDS Res. and Hum. Retroviruses 22, 12, 1220-1230 (2006). – Matzkevich 2006.pdf
- Moelling, K., Matskevich, A. and Jung, J.-S. Relationship between retroviral replication and RNA interference machineries. Cold Spring Harbor Symp. on Quant. Biol., Vol. 71, 365-368 (2006). – Moelling 2007.pdf
- Matzen, K., Elzaouk, L., Matskevich A.A., Nitzsche, A., Heinrich, J. and Moelling, K. RNase H-mediated retrovirus destruction in vivo triggered by oligodeoxynucleotides. Nature Biotechnol. 25, 669-674 (2007). Matzen 2007.pdf Editorial Comment: Assisted suicide for retroviruses Nature Biotechnol. 25, 643-644 (2007) Comment Matzen 2007.pdf
- Heinrich, J., Mathur, S., Matskevich, A.A., and Moelling, K. Oligonucleotide-mediated retroviral RNase H activation leads to reduced HIV-1 titer in patient-derived plasma. AIDS 23, 213-221 (2009). Heinrich 2009.pdf Editorial Comment: Mark Wainberg: New findings on blockage of HIV-1 RNase H activity – Comment Heinrich 2009.pdf
- Wittmer-Elzaouk, L., Jung-Shiu, J., Heinrich, J., and Moelling, K. Retroviral self-inactivation in the mouse vagina induced by short DNA. Antiviral Res 82, 22-28 (2009) – Wittmer-Elzaouk 2009
- Falkenhagen, A., Heinrich, J., and Moelling, K. Short hairpin-loop-structured Oligodeoxynucleotides reduce HSV-1 Replication. Virology Journal 6, 43 (2009).
- Kwok, T., Heinrich, J., Jung-Shiu, J., Meier, M.G., Mathur, S., and Moelling, K. Reduction of gene expression by a hairpin-loop structured oligodeoxynucleotide: alternative to siRNA and antisense. Biochim Biophys. Acta 1790, 1170-8 (2009) – Kwok 2009b
- Hofmann M., Heinrich, J., Radziwill, G., and Moelling, K. A short hairpin DNA analogous to miR125b inhibits C-Raf expression, proliferation and survival in breast cancer cells. Molecular Cancer Research 7, 1635-1644 (2009).