Differential contribution of basic residues to HIV-1 nucleocapsid protein's nucleic acid chaperone function and retroviral replication

Hao Wu, Mithun Mitra, M. Nabuan Naufer, Micah J. McCauley, Robert J. Gorelick, Ioulia Rouzina, Karin Musier-Forsyth, Mark C. Williams

Research output: Contribution to journalArticlepeer-review

24 Scopus citations

Abstract

The human immunodeficiency virus type 1 (HIV-1) nucleocapsid (NC) protein contains 15 basic residues located throughout its 55-amino acid sequence, as well as one aromatic residue in each of its two CCHC-type zinc finger motifs. NC facilitates nucleic acid (NA) rearrangements via its chaperone activity, but the structural basis for this activity and its consequences in vivo are not completely understood. Here, we investigate the role played by basic residues in the N-terminal domain, the N-terminal zinc finger and the linker region between the two zinc fingers. We use in vitro ensemble and single-molecule DNA stretching experiments to measure the characteristics of wild-type and mutant HIV-1 NC proteins, and correlate these results with cell-based HIV-1 replication assays. All of the cationic residue mutations lead to NA interaction defects, as well as reduced HIV-1 infectivity, and these effects are most pronounced on neutralizing all five N-terminal cationic residues. HIV-1 infectivity in cells is correlated most strongly with NC's NA annealing capabilities as well as its ability to intercalate the DNA duplex. Although NC's aromatic residues participate directly in DNA intercalation, our findings suggest that specific basic residues enhance these interactions, resulting in optimal NA chaperone activity.

Original languageEnglish (US)
Pages (from-to)2525-2537
Number of pages13
JournalNucleic acids research
Volume42
Issue number4
DOIs
StatePublished - Feb 2014

Bibliographical note

Funding Information:
Federal funds from the National Cancer Institute, National Institutes of Health, under contract HHSN261200800001E with Leidos Biomedical Research, Inc. (RJG); National Institutes of Health [GM065056 to K.M.-F. and GM072462 to M.C.W.]; National Science Foundation [MCB-1243883 to M.C.W]. Funding for open access charge: National Institutes of Health.

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