Differing Roles of the N- and C-terminal Zinc Fingers in Human Immunodeficiency Virus Nucleocapsid Protein-enhanced Nucleic Acid Annealing

The replication process of human immunodeficiency virus requires a number of nucleic acid annealing steps facilitated by the hybridization and helix-destabilizing activities of human immunodeficiency virus nucleocapsid (NC) protein. NC contains two CCHC zinc finger motifs numbered 1 and 2 from the N...

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Bibliographic Details
Published in:The Journal of biological chemistry 2003-08, Vol.278 (33), p.30755-30763
Main Authors: Heath, Megan J., Derebail, Suchitra S., Gorelick, Robert J., DeStefano, Jeffrey J.
Format: Article
Language:English
Subjects:
DNA, Complementary - chemistry
DNA, Complementary - genetics
Fluorescence Resonance Energy Transfer
HIV-1 - genetics
HIV-1 - growth & development
Mutation
Nucleic Acid Conformation
Nucleocapsid Proteins - chemistry
Nucleocapsid Proteins - genetics
Oligonucleotides - chemistry
Oligonucleotides - genetics
Protein Structure, Tertiary
RNA - chemistry
RNA - genetics
Zinc Fingers - genetics
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Summary:The replication process of human immunodeficiency virus requires a number of nucleic acid annealing steps facilitated by the hybridization and helix-destabilizing activities of human immunodeficiency virus nucleocapsid (NC) protein. NC contains two CCHC zinc finger motifs numbered 1 and 2 from the N terminus. The amino acids surrounding the CCHC residues differ between the two zinc fingers. Assays were preformed to investigate the activities of the fingers by determining the effect of mutant and wild-type proteins on annealing of 42-nucleotide RNA and DNA complements. The mutants 1.1 NC and 2.2 NC had duplications of the N- and C-terminal zinc fingers in positions 1 and 2. The mutant 2.1 NC had the native zinc fingers with their positions switched. Annealing assays were completed with unstructured and highly structured oligonucleotide complements. 2.2 NC had a near wild-type level of annealing of unstructured nucleic acids, whereas it was completely unable to stimulate annealing of highly structured nucleic acids. In contrast, 1.1 NC was able to stimulate annealing of both unstructured and structured substrates, but to a lesser degree than the wild-type protein. Results suggest that finger 1 has a greater role in unfolding of strong secondary structures, whereas finger 2 serves an accessory role that leads to a further increase in the rate of annealing.
ISSN:0021-9258
1083-351X
DOI:10.1074/jbc.M303819200