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 Rothenberg, S. M.
Right arrow Articles by Brown, P. O.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Rothenberg, S. M.
Right arrow Articles by Brown, P. O.

 Previous Article  |  Next Article 

Journal of Virology, December 2001, p. 11851-11862, Vol. 75, No. 23
0022-538X/01/$04.00+0   DOI: 10.1128/JVI.75.23.11851-11862.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.

Comprehensive Mutational Analysis of the Moloney Murine Leukemia Virus Envelope Protein

S. Michael Rothenberg,1,2 Mari N. Olsen,1,3 Louise Chang Laurent,4,dagger Rachel Adams Crowley,1,2 and Patrick O. Brown1,3,*

Department of Biochemistry,2 Program in Cancer Biology,1 and Howard Hughes Medical Institute,3 Stanford University Medical Center, Palo Alto, California 94305, and Department of Biochemistry, University of California at San Francisco, San Francisco, California 941434

Received 20 June 2001/Accepted 23 August 2001

The envelope (Env) protein of Moloney murine leukemia virus is the primary mediator of viral entry. We constructed a large pool of insertion mutations in the env gene and analyzed the fitness of each mutant in completing two critical steps in the virus life cycle: (i) the expression and delivery of the Env protein to the cell surface during virion assembly and (ii) the infectivity of virions displaying the mutant proteins. The majority of the mutants were poorly expressed at the producer cell surface, suggesting folding defects due to the presence of the inserted residues. The mutants with residual infectivity had insertions either in the amino-terminal signal sequence region, two disulfide-bonded loops in the receptor binding domain, discrete regions of the carboxy-terminal region of the surface subunit (SU), or the cytoplasmic tail. Insertions that allowed the mutants to reach the cell surface but not to mediate detectable infection were located within the amino-terminal sequence of the mature Env, within the SU carboxy-terminal region, near putative receptor binding residues, and throughout the fusion peptide. Independent analysis of select mutants in this group allowed more precise identification of the defect in Env function. Mapping of mutant phenotypes to a structural model of the receptor-binding domain provides insights into the protein's functional organization. The high-resolution functional map reported here will be valuable for the engineering of the Env protein for a variety of uses, including gene therapy.

* Corresponding author. Mailing address: Howard Hughes Medical Institute, Stanford University Medical Center, 279 Campus Dr., Beckman Center B439, Palo Alto, CA 94305. Phone: (650) 725-7567. Fax: (650) 723-1399. E-mail: pbrown{at}cmgm.stanford.edu.

dagger Present address: University of California at San Diego, Department of Reproductive Medicine, Division of Obstetrics and Gynecology, San Diego, CA 92103.

Journal of Virology, December 2001, p. 11851-11862, Vol. 75, No. 23
0022-538X/01/$04.00+0   DOI: 10.1128/JVI.75.23.11851-11862.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.


This article has been cited by other articles:

  • Pajunen, M., Turakainen, H., Poussu, E., Peranen, J., Vihinen, M., Savilahti, H. (2007). High-precision mapping of protein protein interfaces: an integrated genetic strategy combining en masse mutagenesis and DNA-level parallel analysis on a yeast two-hybrid platform. Nucleic Acids Res 0: gkm563v1-11 [Abstract] [Full Text]  
  • Ou, W., Silver, J. (2006). Stoichiometry of Murine Leukemia Virus Envelope Protein-Mediated Fusion and Its Neutralization. J. Virol. 80: 11982-11990 [Abstract] [Full Text]  
  • Puglia, J., Wang, T., Smith-Snyder, C., Cote, M., Scher, M., Pelletier, J. N., John, S., Jonsson, C. B., Roth, M. J. (2006). Revealing Domain Structure through Linker-Scanning Analysis of the Murine Leukemia Virus (MuLV) RNase H and MuLV and Human Immunodeficiency Virus Type 1 Integrase Proteins. J. Virol. 80: 9497-9510 [Abstract] [Full Text]  
  • Klymiuk, N., Muller, M., Brem, G., Aigner, B. (2006). Phylogeny, recombination and expression of porcine endogenous retrovirus {gamma}2 nucleotide sequences.. J. Gen. Virol. 87: 977-986 [Abstract] [Full Text]  
  • Ou, W., Lu, N., Yu, S. S., Silver, J. (2006). Effect of Epitope Position on Neutralization by Anti-Human Immunodeficiency Virus Monoclonal Antibody 2F5. J. Virol. 80: 2539-2547 [Abstract] [Full Text]  
  • Burkhart, M. D., D'Agostino, P., Kayman, S. C., Pinter, A. (2005). Involvement of the C-Terminal Disulfide-Bonded Loop of Murine Leukemia Virus SU Protein in a Postbinding Step Critical for Viral Entry. J. Virol. 79: 7868-7876 [Abstract] [Full Text]  
  • Ye, K., Jin, S., Ataai, M. M., Schultz, J. S., Ibeh, J. (2004). Tagging Retrovirus Vectors with a Metal Binding Peptide and One-Step Purification by Immobilized Metal Affinity Chromatography. J. Virol. 78: 9820-9827 [Abstract] [Full Text]  
  • Bubeck, A., Wagner, M., Ruzsics, Z., Lotzerich, M., Iglesias, M., Singh, I. R., Koszinowski, U. H. (2004). Comprehensive Mutational Analysis of a Herpesvirus Gene in the Viral Genome Context Reveals a Region Essential for Virus Replication. J. Virol. 78: 8026-8035 [Abstract] [Full Text]  
  • Moradpour, D., Evans, M. J., Gosert, R., Yuan, Z., Blum, H. E., Goff, S. P., Lindenbach, B. D., Rice, C. M. (2004). Insertion of Green Fluorescent Protein into Nonstructural Protein 5A Allows Direct Visualization of Functional Hepatitis C Virus Replication Complexes. J. Virol. 78: 7400-7409 [Abstract] [Full Text]  
  • O'Reilly, L., Roth, M. J. (2003). G541R within the 4070A TM Protein Regulates Fusion in Murine Leukemia Viruses. J. Virol. 77: 12011-12021 [Abstract] [Full Text]  
  • Auerbach, M. R., Shu, C., Kaplan, A., Singh, I. R. (2003). Functional characterization of a portion of the Moloney murine leukemia virus gag gene by genetic footprinting. Proc. Natl. Acad. Sci. USA 100: 11678-11683 [Abstract] [Full Text]  
  • Kubo, Y., Amanuma, H. (2003). Mutational analysis of the R peptide cleavage site of Moloney murine leukaemia virus envelope protein. J. Gen. Virol. 84: 2253-2257 [Abstract] [Full Text]  
  • LAND, A., ZONNEVELD, D., BRAAKMAN, I. (2003). Folding of HIV-1 Envelope glycoprotein involves extensive isomerization of disulfide bonds and conformation-dependent leader peptide cleavage. FASEB J. 17: 1058-1067 [Abstract] [Full Text]  
  • Lu, C.-W., O'Reilly, L., Roth, M. J. (2002). G100R Mutation within 4070A Murine Leukemia Virus Env Increases Virus Receptor Binding, Kinetics of Entry, and Viral Transduction Efficiency. J. Virol. 77: 739-743 [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»