Nucleic acid binding and chaperone properties of HIV-1 Gag and nucleocapsid proteins.
The Gag polyprotein of HIV-1 is essential for retroviral replication and packaging. The nucleocapsid (NC) protein is the primary region for the interaction of Gag with nucleic acids. In this study, we examine the interactions of Gag and its NC cleavage products (NCp15, NCp9 and NCp7) with nucleic acids using solution and single molecule experiments. The NC cleavage products bound DNA with comparable affinity and strongly destabilized the DNA duplex. In contrast, the binding constant of Gag to DNA was found to be approximately 10-fold higher than that of the NC proteins, and its destabilizing effect on dsDNA was negligible. These findings are consistent with the primary function of Gag as a nucleic acid binding and packaging protein and the primary function of the NC proteins as nucleic acid chaperones. Also, our results suggest that NCp7's capability for fast sequence-nonspecific nucleic acid duplex destabilization, as well as its ability to facilitate nucleic acid strand annealing by inducing electrostatic attraction between strands, likely optimize the fully processed NC protein to facilitate complex nucleic acid secondary structure rearrangements. In contrast, Gag's stronger DNA binding and aggregation capabilities likely make it an effective chaperone for processes that do not require significant duplex destabilization.
Cruceanu
Margareta
M
Department of Physics, Northeastern University, 111 Dana Research Center, 110 Forsyth Street, Boston, MA 02115, USA.
Urbaneja
Maria A
MA
Hixson
Catherine V
CV
Johnson
Donald G
DG
Datta
Siddhartha A
SA
Fivash
Matthew J
MJ
Stephen
Andrew G
AG
Fisher
Robert J
RJ
Gorelick
Robert J
RJ
Casas-Finet
Jose R
JR
Rein
Alan
A
Rouzina
Ioulia
I
Williams
Mark C
MC
D036022:Capsid Proteins / Q000378:metabolism*; D004247:DNA / Q000737:chemistry / Q000378:metabolism*; D005454:Fluorescence Polarization; D015683:Gene Products, gag / Q000378:metabolism*; D015497:HIV-1 / Q000502:physiology*; D018832:Molecular Chaperones / Q000378:metabolism*; D009690:Nucleic Acid Conformation; D009691:Nucleic Acid Denaturation; D019590:Nucleocapsid Proteins / Q000378:metabolism*; D011498:Protein Precursors / Q000378:metabolism; D012313:RNA / Q000378:metabolism; D013050:Spectrometry, Fluorescence; D014764:Viral Proteins / Q000378:metabolism*; D019065:Virus Assembly; D054301:gag Gene Products, Human Immunodeficiency Virus
D036022:Capsid Proteins; D015683:Gene Products, gag; D018832:Molecular Chaperones; C071884:NCP7 protein, Human immunodeficiency virus 1; D019590:Nucleocapsid Proteins; D011498:Protein Precursors; D014764:Viral Proteins; D054301:gag Gene Products, Human Immunodeficiency Virus; C060295:p15 gag protein, Human immunodeficiency virus 1; D012313:RNA; D004247:DNA