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Journal of Virology, May 2002, p. 4950-4960, Vol. 76, No. 10
0022-538X/02/$04.00+0     DOI: 10.1128/JVI.76.10.4950-4960.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.

cis-Acting Elements Important for Retroviral RNA Packaging Specificity

Benjamin E. Beasley1,2 and Wei-Shau Hu1*

HIV Drug Resistance Program, National Cancer Institute, Frederick, Maryland 21702,1 Department of Microbiology, Immunology, and Cell Biology, School of Medicine, West Virginia University, Morgantown, West Virginia 265062

Received 12 November 2001/ Accepted 20 February 2002

Spleen necrosis virus (SNV) proteins can package RNA from distantly related murine leukemia virus (MLV), whereas MLV proteins cannot package SNV RNA efficiently. We used this nonreciprocal recognition to investigate regions of packaging signals that influence viral RNA encapsidation specificity. Although the MLV and SNV packaging signals ({Psi} and E, respectively) do not contain significant sequence homology, they both contain a pair of hairpins. This hairpin pair was previously proposed to be the core element in MLV {Psi}. In the present study, MLV-based vectors were generated to contain chimeric SNV/MLV packaging signals in which the hairpins were replaced with the heterologous counterpart. The interactions between these chimeras and MLV or SNV proteins were examined by virus replication and RNA analyses. SNV proteins recognized all of the chimeras, indicating that these chimeras were functional. We found that replacing the hairpin pair did not drastically alter the ability of MLV proteins to package these chimeras. These results indicate that, despite the important role of the hairpin pair in RNA packaging, it is not the major motif responsible for the ability of MLV proteins to discriminate between the MLV and SNV packaging signals. To determine the role of sequences flanking the hairpins in RNA packaging specificity, vectors with swapped flanking regions were generated and evaluated. SNV proteins packaged all of these chimeras efficiently. In contrast, MLV proteins strongly favored chimeras with the MLV 5'-flanking regions. These data indicated that MLV Gag recognizes multiple elements in the viral packaging signal, including the hairpin structure and flanking regions.

* Corresponding author. Mailing address: HIV Drug Resistance Program, National Cancer Institute, NCI at Frederick, P.O. Box B, Building 535, Room 336, Frederick, MD 21702-1201. Phone: (301) 846-1250. Fax: (301) 846-6013. E-mail: whu{at}ncifcrf.gov.

Journal of Virology, May 2002, p. 4950-4960, Vol. 76, No. 10
0022-538X/02/$04.00+0     DOI: 10.1128/JVI.76.10.4950-4960.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.



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