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G-quartets direct assembly of HIV-1 nucleocapsid protein along single-stranded DNA

  1. Author:
    Lyonnais, S.
    Gorelick, R. J.
    Mergny, J. L.
    Le Cam, E.
    Mirambeau, G.

  2. Author Address

    Inst Gustave Roussy, CNRS, UMR 8126, Lab Microscopie Mol & Cellulaire, 39 Rue Camille Desmoulins, F-94805 Villejuif, France Inst Gustave Roussy, CNRS, UMR 8126, Lab Microscopie Mol & Cellulaire, F-94805 Villejuif, France NCI, AIDS Vaccine Programe, SAIC Frederick Inc, Frederick, MD 21702 USA CNRS, INSERM, UMR 8646,U565, Museum Natl Hist Nat,Lab Biophys, F-75231 Paris, France Mirambeau G Inst Gustave Roussy, CNRS, UMR 8126, Lab Microscopie Mol & Cellulaire, 39 Rue Camille Desmoulins, F-94805 Villejuif, France
    1. Year: 2003
  2. Journal: Nucleic Acids Research
    1. 31
    2. 19
    3. Pages: 5754-5763
  4. Type of Article: Article
  1. Abstract:

    The d(TTGGGGGGTACAGTGCA) sequence, derived from the human immunodeficiency virus type 1 (HIV-1) central DNA flap, can form in vitro an intermolecular parallel DNA quadruplex. This work demonstrates that the HIV-1 nucleocapsid protein (NCp) exhibits a high affinity (10(8) M-1) for this quadruplex. This interaction is predominantly hydrophobic, maintained by a stabilization between G-quartet planes and the C-terminal zinc finger of the protein. It also requires 5 nt long tails flanking the quartets plus both the second zinc-finger and the N-terminal domain of NCp. The initial binding nucleates an ordered arrangement of consecutive NCp along the four single- stranded tails. Such a process requires the N-terminal zinc finger, and was found to occur for DNA site sizes shorter than usual in a sequence-dependent manner. Concurrently, NCp binding is efficient on a G'2 quadruplex also derived from the HIV-1 central DNA flap. Apart from their implication within the DNA flap, these data lead to a model for the nucleic acid architecture within the viral nucleocapsid, where adjacent single-stranded tails and NCp promote a compact assembly of NCp and nucleic acid growing from stably and primary bound NCp.

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