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Journal of Virology, July 2004, p. 7677-7684, Vol. 78, No. 14
0022-538X/04/$08.00+0     DOI: 10.1128/JVI.78.14.7677-7684.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.

RNA Sequences in the Moloney Murine Leukemia Virus Genome Bound by the Gag Precursor Protein in the Yeast Three-Hybrid System

Matthew J. Evans,1,{dagger} Eran Bacharach,2,{ddagger} and Stephen P. Goff1,2*

Integrated Program in Cellular, Molecular, and Biophysical Studies,1 Howard Hughes Medical Institute, Department of Biochemistry and Biophysics, College of Physicians and Surgeons, Columbia University, New York, New York 100322

Received 26 January 2004/ Accepted 6 April 2004

Encapsidation of the Moloney murine leukemia virus (MMLV) genome is mediated through a specific interaction between the major viral structural protein, Gag, and an RNA packaging signal, {Psi}. Many studies have investigated this process in vivo, although the specific examination of the Gag-RNA interaction in this context is difficult due to the variety of other viral functions involved in virion assembly in vivo. The Saccharomyces cerevisiae three-hybrid assay was used to directly examine the interaction between MMLV Gag and {Psi}. In this system, MMLV RNA regions exhibiting high-affinity Gag binding were mapped. All Gag-binding regions were located 3' to the viral splice donor sequence of the viral RNA transcript. No single short RNA sequence within {Psi} supported strong Gag interaction. Instead, an RNA comprised of nearly the entire {Psi} region was necessary to demonstrate an appreciable Gag interaction in the yeast three-hybrid system. These finding support the notion that two stem-loops (C and D) are not sufficient to form a core MMLV encapsidation signal.

* Corresponding author. Mailing address: Department of Biochemistry and Biophysics, College of Physicians and Surgeons, Columbia University, New York, NY 10032. Phone: (212) 305-3794. Fax: (212) 305-8692. E-mail: goff{at}cancercenter.columbia.edu.

{dagger} Present address: Center for the Study of Hepatitis C, Laboratory of Virology and Infectious Disease, The Rockefeller University, New York, NY 10021.

{ddagger} Present address: Department of Cell Research and Immunology, Tel Aviv University, Tel Aviv, 69978 Israel.

Journal of Virology, July 2004, p. 7677-7684, Vol. 78, No. 14
0022-538X/04/$08.00+0     DOI: 10.1128/JVI.78.14.7677-7684.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.



This article has been cited by other articles:

  • Gherghe, C., Weeks, K. M. (2006). The SL1-SL2 (Stem-Loop) Domain Is the Primary Determinant for Stability of the Gamma Retroviral Genomic RNA Dimer. J. Biol. Chem. 281: 37952-37961 [Abstract] [Full Text]  
  • Mark-Danieli, M., Laham, N., Kenan-Eichler, M., Castiel, A., Melamed, D., Landau, M., Bouvier, N. M., Evans, M. J., Bacharach, E. (2005). Single Point Mutations in the Zinc Finger Motifs of the Human Immunodeficiency Virus Type 1 Nucleocapsid Alter RNA Binding Specificities of the Gag Protein and Enhance Packaging and Infectivity. J. Virol. 79: 7756-7767 [Abstract] [Full Text]  


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