HIV / RNase H

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

  1. 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).
  2. Moelling, K.: Characterization of Reverse Transcriptase and RNase H from Friend murine leukemia virus. Virology 62, 46-59 (1974b).
  3. 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).
  4. Moelling, K. Exciting days over for reverse transcriptase? Nature 256, 258-260 (1975).
  5. Moelling, K. Comparison between an avian and a murine viral reverse transcriptase RNase H complex. Comparative Leukemia Res., Bibl. Haematol. 43, 121-124 (1976).
  6. Moelling, K. Further characterization of the Friend murine leukemia virus RNase H complex. J. Virol. 18, 418-425 (1976).
  7. Moelling, K. Anything new about reverse transcriptase? Nature 264, 111-112 (1976)
  8. 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).
  9. 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).
  10. Sykora, K.W. and Moelling, K. Properties of the avian viral protein p12. J. Gen. Virol. 55, 379-391 (1981).
  11. 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).
  12. 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
  13. Moelling, K., Schulze, T. and Diringer, H. Inhibition of HIV-1 RNase H by sulfated polyanions. J. Virol. 63, 5489-5492 (1989).
  14. 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
  15. Wöhrl, B.M., Moelling, K. Coupling of reverse transcriptase and RNase H during HIV-1 replication. Behring Inst Mitt. (1991)
  16. 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
  17. 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).
  18. 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).
  19. 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).
  20. 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).
  21. 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
  22. 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
  23. 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
  24. 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
  25. 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
  26. 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
  27. 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
  28. 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
  29. 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
  30. Falkenhagen, A., Heinrich, J., and Moelling, K. Short hairpin-loop-structured Oligodeoxynucleotides reduce HSV-1 Replication. Virology Journal 6, 43 (2009).
  31. 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
  32. 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).