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J Virol, March 1998, p. 1902-1909, Vol. 72, No. 3
0022-538X/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.

Sequence-Specific Binding of Human Immunodeficiency Virus Type 1 Nucleocapsid Protein to Short Oligonucleotides

Robert J. Fisher,1,* Alan Rein,2 Matthew Fivash,3 Maria A. Urbaneja,4 José R. Casas-Finet,4 Maxine Medaglia,1 and Louis E. Henderson4

Protein Chemistry Laboratory1 and AIDS Vaccine Development Program,4 SAIC Frederick, Retroviral Genetics Section, ABL-Basic Research Program,2 and Data Management Services,3 NCI-Frederick Cancer Research and Development Center, Frederick, Maryland 21702

Received 31 July 1997/Accepted 1 December 1997

We have analyzed the binding of recombinant human immunodeficiency virus type 1 nucleocapsid protein (NC) to very short oligonucleotides by using surface plasmon resonance (SPR) technology. Our experiments, which were conducted at a moderate salt concentration (0.15 M NaCl), showed that NC binds more stably to runs of d(G) than to other DNA homopolymers. However, it exhibits far more stable binding with the alternating base sequence d(TG)n than with any homopolymeric oligodeoxyribonucleotide; thus, it shows a strong sequence preference under our experimental conditions. We found that the minimum length of an alternating d(TG) sequence required for stable binding was five nucleotides. Stable binding to the tetranucleotide d(TG)2 was observed only under conditions where two tetranucleotide molecules were held in close spatial proximity. The stable, sequence-specific binding to d(TG)n required that both zinc fingers be present, each in its proper position in the NC protein, and was quite salt resistant, indicating a large hydrophobic contribution to the binding. Limited tests with RNA oligonucleotides indicated that the preferential sequence-specific binding observed with DNA also occurs with RNA. Evidence was also obtained that NC can bind to nucleic acid molecules in at least two distinct modes. The biological significance of the specific binding we have detected is not known; it may reflect the specificity with which the parent Gag polyprotein packages genomic RNA or may relate to the functions of NC after cleavage of the polyprotein, including its role as a nucleic acid chaperone.

* Corresponding author. Mailing address: SAIC, Frederick, NCI-Frederick Cancer Research and Development Center, P.O. Box B, Bldg. 469, Frederick, MD 21702. Phone: (301) 846-1633. Fax: (301) 846-6065. E-mail: fisher{at}ncifcrf.gov.




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