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Journal of Virology, January 2000, p. 541-546, Vol. 74, No. 1
0022-538X/0/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.

A Heterologous, High-Affinity RNA Ligand for Human Immunodeficiency Virus Gag Protein Has RNA Packaging Activity

Jared L. Clever,1,2,dagger Randy A. Taplitz,3,Dagger Michael A. Lochrie,4 Barry Polisky,4,§ and Tristram G. Parslow1,2,*

Departments of Pathology,1 Microbiology and Immunology,2 and Internal Medicine,3 University of California, San Francisco, California 94143-0506, and NeXstar Pharmaceuticals, Inc., Boulder, Colorado 803014

Received 22 July 1999/Accepted 28 September 1999

Retroviral RNA encapsidation depends on the specific binding of Gag proteins to packaging (psi ) signals in genomic RNA. We investigated whether an in vitro-selected, high-affinity RNA ligand for the nucleocapsid (NC) portion of the Gag protein from human immunodeficiency virus type 1 (HIV-1) could mediate packaging into HIV-1 virions. We find that this ligand can functionally substitute for one of the Gag-binding elements (termed SL3) in the HIV-1 psi  locus to support packaging and viral infectivity in cis. By contrast, this ligand, which fails to dimerize spontaneously in vitro, is unable to replace a different psi  element (termed SL1) which is required for both Gag binding and dimerization of the HIV-1 genome. A single point mutation within the ligand that eliminates high-affinity in vitro Gag binding also abolishes its packaging activity at the SL3 position. These results demonstrate that specific binding of Gag or NC protein is a critical determinant of genomic RNA packaging.

* Corresponding author. Mailing address: Department of Pathology, Box 0506, University of California, San Francisco, CA 94143-0506. Phone: (415) 476-1015. Fax: (415) 476-9672. E-mail: parslow{at}cgl.ucsf.edu.

dagger Present address: Department of Microbiology, University of Texas Health Sciences Center at San Antonio, San Antonio, TX 78284-7758.

Dagger Present address: Department of Medicine, Oregon Health Sciences University, Portland, OR 97201.

§ Present address: BioStar, Inc., Boulder, CO 80301.

Journal of Virology, January 2000, p. 541-546, Vol. 74, No. 1
0022-538X/0/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.


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