This Article
Right arrow Full Text
Right arrow Full Text (PDF)
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrowReprints and Permissions
Right arrow Copyright Information
Right arrow Books from ASM Press
Right arrow MicrobeWorld
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Pfrepper, K.-I.
Right arrow Articles by Flügel, R. M.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Pfrepper, K.-I.
Right arrow Articles by Flügel, R. M.

 Previous Article  |  Next Article 

Journal of Virology, September 1998, p. 7648-7652, Vol. 72, No. 9
0022-538X/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.

Molecular Characterization of Proteolytic Processing of the Pol Proteins of Human Foamy Virus Reveals Novel Features of the Viral Protease

Klaus-Ingmar Pfrepper,1 Hans-Richard Rackwitz,2 Martina Schnölzer,3 Hans Heid,2 Martin Löchelt,1 and Rolf M. Flügel1,*

Abteilungen Retroviral Gene Expression, Research Program Applied Tumorvirology,1 Cell Biology, Research Program Cell Differentiation and Carcinogenesis,2 and Central Protein Analysis Group,3 German Cancer Research Center, 69009 Heidelberg, Federal Republic of Germany

Received 6 February 1998/Accepted 5 May 1998

Spumaviruses, or foamy viruses, express a pol-specific transcript that codes for a Pol polyprotein that consists of the protease, reverse transcriptase, ribonuclease H, and the integrase domains. To delineate the proteolytic cleavage sites between the Pol subdomains, recombinant human foamy virus (HFV) Pol proteins were expressed, purified by affinity chromatography, and subjected to either HFV protease assays or autocatalytic processing. In control experiments, HFV protease-deficient mutant proteins in which the active site Asp was replaced by an Ala residue were used to rule out unspecific processing by nonviral proteases. Specific proteolytic cleavage products were isolated, and the cleavage sites were analyzed by amino acid sequencing. Peptides spanning the resulting cleavage sites were chemically synthesized and assayed with HFV protease, and the cleaved peptides were subjected to mass spectrometry. The cleavage site sequences obtained were in complete agreement with the amino-terminal sequences from amino acid sequencing of authentic cleavage products of the HFV Pol proteins. Analysis by fast-protein liquid chromatography of a short version of the active HFV protease revealed that the enzyme predominantly formed dimeric molecules.

* Corresponding author. Mailing address: Abteilung Retroviral Gene Expression, Applied Tumorvirology, DKFZ, INF 242, 69009 Heidelberg, Germany. Phone: 49-6221-424611. Fax: 49-6221-424865. E-mail: r.m.fluegel{at}dkfz-heidelberg.de.

Journal of Virology, September 1998, p. 7648-7652, Vol. 72, No. 9
0022-538X/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.


This article has been cited by other articles:

  • Valkov, E., Gupta, S. S., Hare, S., Helander, A., Roversi, P., McClure, M., Cherepanov, P. (2009). Functional and structural characterization of the integrase from the prototype foamy virus. Nucleic Acids Res 37: 243-255 [Abstract] [Full Text]  
  • Hartl, M. J., Kretzschmar, B., Frohn, A., Nowrouzi, A., Rethwilm, A., Wohrl, B. M. (2008). AZT resistance of simian foamy virus reverse transcriptase is based on the excision of AZTMP in the presence of ATP. Nucleic Acids Res 36: 1009-1016 [Abstract] [Full Text]  
  • Roy, J., Linial, M. L. (2007). Role of the Foamy Virus Pol Cleavage Site in Viral Replication. J. Virol. 81: 4956-4962 [Abstract] [Full Text]  
  • Sperka, T., Boross, P., Eizert, H., Tozser, J., Bagossi, P. (2006). Effect of mutations on the dimer stability and the pH optimum of the human foamy virus protease. Protein Eng Des Sel 19: 369-375 [Abstract] [Full Text]  
  • Abram, M. E., Parniak, M. A. (2005). Virion Instability of Human Immunodeficiency Virus Type 1 Reverse Transcriptase (RT) Mutated in the Protease Cleavage Site between RT p51 and the RT RNase H Domain. J. Virol. 79: 11952-11961 [Abstract] [Full Text]  
  • Boyer, P. L., Stenbak, C. R., Clark, P. K., Linial, M. L., Hughes, S. H. (2004). Characterization of the Polymerase and RNase H Activities of Human Foamy Virus Reverse Transcriptase. J. Virol. 78: 6112-6121 [Abstract] [Full Text]  
  • Verschoor, E. J., Langenhuijzen, S., van den Engel, S., Niphuis, H., Warren, K. S., Heeney, J. L. (2003). Structural and Evolutionary Analysis of an Orangutan Foamy Virus. J. Virol. 77: 8584-8587 [Abstract] [Full Text]  
  • Rinke, C. S., Boyer, P. L., Sullivan, M. D., Hughes, S. H., Linial, M. L. (2002). Mutation of the Catalytic Domain of the Foamy Virus Reverse Transcriptase Leads to Loss of Processivity and Infectivity. J. Virol. 76: 7560-7570 [Abstract] [Full Text]  
  • Yvon-Groussin, A., Mugnier, P., Bertin, P., Grandadam, M., Agut, H., Huraux, J. M., Calvez, V. (2001). Efficacy of Dideoxynucleosides against Human Foamy Virus and Relationship to Its Reverse Transcriptase Amino Acid Sequence and Structure. J. Virol. 75: 7184-7187 [Abstract] [Full Text]  
  • Imrich, H., Heinkelein, M., Herchenröder, O., Rethwilm, A. (2000). Primate foamy virus Pol proteins are imported into the nucleus. J. Gen. Virol. 81: 2941-2947 [Abstract] [Full Text]  
  • Tobaly-Tapiero, J., Bittoun, P., Neves, M., Guillemin, M.-C., Lecellier, C.-H., Puvion-Dutilleul, F., Gicquel, B., Zientara, S., Giron, M.-L., de Thé, H., Saïb, A. (2000). Isolation and Characterization of an Equine Foamy Virus. J. Virol. 74: 4064-4073 [Abstract] [Full Text]  
  • Pfrepper, K.-I., Löchelt, M., Rackwitz, H.-R., Schnölzer, M., Heid, H., Flügel, R. M. (1999). Molecular Characterization of Proteolytic Processing of the Gag Proteins of Human Spumavirus. J. Virol. 73: 7907-7911 [Abstract] [Full Text]  
  • Baldwin, D. N., Linial, M. L. (1999). Proteolytic Activity, the Carboxy Terminus of Gag, and the Primer Binding Site Are Not Required for Pol Incorporation into Foamy Virus Particles. J. Virol. 73: 6387-6393 [Abstract] [Full Text]  
  • Linial, M. L. (1999). Foamy Viruses Are Unconventional Retroviruses. J. Virol. 73: 1747-1755 [Full Text]  
  • Mulder, L. C. F., Muesing, M. A. (2000). Degradation of HIV-1 Integrase by the N-end Rule Pathway. J. Biol. Chem. 275: 29749-29753 [Abstract] [Full Text]  


Copyright © 2009 by the American Society for Microbiology.   For an alternate route to Journals.ASM.org, visit: http://intl-journals.asm.org | More Info»