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Journal of Virology, January 2005, p. 834-840, Vol. 79, No. 2
0022-538X/05/$08.00+0     doi:10.1128/JVI.79.2.834-840.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.

Directed Evolution of Retrovirus Envelope Protein Cytoplasmic Tails Guided by Functional Incorporation into Lentivirus Particles

Christoph A. Merten,1 Jörn Stitz,2 Gundula Braun,1 Eric M. Poeschla,3 Klaus Cichutek,1 and Christian J. Buchholz1*

Division of Medical Biotechnology, Paul-Ehrlich-Institut, Langen, Germany,1 Department of Molecular Pharmacology, School of Medicine, Stanford University, Stanford, California,2 Molecular Medicine Program, Departments of Immunology and Medicine, Mayo Medical School, Rochester, Minnesota3

Received 3 March 2004/ Accepted 3 September 2004

In contrast to most gammaretrovirus envelope proteins (Env), the Gibbon ape leukemia virus (GaLV) Env protein does not mediate the infectivity of human immunodeficiency virus type 1 (HIV-1) particles. We made use of this observation to set up a directed evolution system by creating a library of GaLV Env variants diversified at three critical amino acids, all located around the R-peptide cleavage site within the cytoplasmic tail. This library was screened for variants that were able to functionally pseudotype HIV-1 vector particles. All selected Env variants mediated the infectivity of HIV-1 vector particles and encoded novel cytoplasmic tail motifs. They were efficiently incorporated into HIV particles, and the R peptide was processed by the HIV protease. Interestingly, in some of the selected variants, the R-peptide cleavage site had shifted closer to the C terminus. These data demonstrate a valuable approach for the engineering of chimeric viruses and vector particles.

* Corresponding author. Mailing address: Division of Medical Biotechnology, Paul-Ehrlich-Institut, Paul-Ehrlich-Str. 51-59, 63225 Langen, Germany. Phone: 49-6103-77-4011. Fax: 49-6103-77-1255. E-mail: bucch{at}pei.de.

Journal of Virology, January 2005, p. 834-840, Vol. 79, No. 2
0022-538X/05/$08.00+0     doi:10.1128/JVI.79.2.834-840.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.



This article has been cited by other articles:

  • Stitz, J., Wolfrum, N., Buchholz, C. J., Cichutek, K. (2006). Envelope proteins of spleen necrosis virus form infectious human immunodeficiency virus type 1 pseudotype vector particles, but fail to incorporate upon substitution of the cytoplasmic domain with that of Gibbon ape leukemia virus. J. Gen. Virol. 87: 1577-1581 [Abstract] [Full Text]  
  • Merten, C. A., Stitz, J., Braun, G., Medvedovska, J., Cichutek, K., Buchholz, C. J. (2006). Fusoselect: cell-cell fusion activity engineered by directed evolution of a retroviral glycoprotein. Nucleic Acids Res 34: e41-e41 [Abstract] [Full Text]  


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