Subtle alterations of the native zinc finger structures have dramatic effects on the nucleic acid chaperone activity of human immunodeficiency virus type 1 nucleocapsid protein

Journal of Virology
Jianhui GuoJudith G Levin

Abstract

The nucleocapsid protein (NC) of human immunodeficiency virus type 1 has two zinc fingers, each containing the invariant CCHC zinc-binding motif; however, the surrounding amino acid context is not identical in the two fingers. Recently, we demonstrated that zinc coordination is required when NC unfolds complex secondary structures in RNA and DNA minus- and plus-strand transfer intermediates; this property of NC reflects its nucleic acid chaperone activity. Here we have analyzed the chaperone activities of mutants having substitutions of alternative zinc-coordinating residues, i.e., CCHH or CCCC, for the wild-type CCHC motif. We also investigated the activities of mutants that retain the CCHC motifs but have mutations that exchange or duplicate the zinc fingers (mutants 1-1, 2-1, and 2-2); these changes affect amino acid context. Our results indicate that in general, for optimal activity in an assay that measures stimulation of minus-strand transfer and inhibition of nonspecific self-priming, the CCHC motif in the zinc fingers cannot be replaced by CCHH or CCCC and the amino acid context of the fingers must be conserved. Context changes also reduce the ability of NC to facilitate primer removal in plus-strand transfer. In additi...Continue Reading

References

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Citations

Aug 19, 2011·AIDS Research and Human Retroviruses·Abdul A Waheed, Eric O Freed
Aug 9, 2003·Nucleic Acids Research·Nick LeeKarin Musier-Forsyth
Sep 23, 2003·Nucleic Acids Research·Sébastien LyonnaisGilles Mirambeau
Aug 4, 2005·Nucleic Acids Research·Besik I KankiaKarin Musier-Forsyth
Jan 26, 2006·Nucleic Acids Research·Robert J FisherAlan Rein
Feb 2, 2006·Nucleic Acids Research·Margareta CruceanuMark C Williams
Oct 26, 2007·Nucleic Acids Research·Chunjuan TianXiao-Fang Yu
Jun 27, 2002·Proceedings of the National Academy of Sciences of the United States of America·Mark C WilliamsKarin Musier-Forsyth
Jun 26, 2010·Physics of Life Reviews·Kathy R ChaurasiyaMark C Williams
Feb 19, 2008·Virus Research·Vandana Purohit BasuRobert A Bambara
Mar 23, 2004·Journal of Molecular Biology·Michele R S HargittaiKarin Musier-Forsyth
Oct 13, 2006·Analytical Biochemistry·Margareta CruceanuMark C Williams
Oct 31, 2006·Journal of Molecular Biology·Reshma M Anthony, Jeffrey J Destefano
Sep 12, 2006·Journal of Molecular Biology·My-Nuong VoKarin Musier-Forsyth
Oct 12, 2005·Journal of Molecular Biology·Mark Nils HansonRobert A Bambara
Feb 19, 2013·Virus Research·Judith G Levin
Aug 5, 2008·Virus Research·Sonia ZúñigaLuis Enjuanes

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