RNA recognition mechanism of the minimal active domain of the human immunodeficiency virus type-2 nucleocapsid protein

Takashi Matsui, Yoshio Kodera, Hiroshi Endoh, Emi Miyauchi, Hiroyoshi Komatsu, Kazuki Sato, Takeshi Tanaka, Toshiyuki Kohno, Tadakazu Maeda

    Research output: Contribution to journalArticlepeer-review

    8 Citations (Scopus)

    Abstract

    NCp8 of HIV-2 contains two CCHC-type zinc fingers connected by a linker, and is involved in many critical steps of the virus life cycle. It was previously shown that the first zinc finger flanked by the linker is the minimal active domain for specific binding to viral RNA. In our previous study, we determined the three-dimensional structure of NCp8-f1, including the minimal active domain, and found that a hydrogen bond between Asn11 N δH and Arg27 O stabilized the conformation of the linker in the vicinity of the zinc finger [Kodera et al. (1998) Biochemistry 37, 17704-17713]. In this study, RNA binding activities of NCp8-f1 and three types of its mutant peptides were analysed by native PAGE assay. The activity and three-dimensional structure of NCp8-f1/N11A, in which alanine is substituted for Asn11 thereby affecting the conformation of the linker, was analyzed and compared with those of NCp8-f1. We demonstrated that the existence of Arg4 and/or Lys5 and Arg26 and/or Arg 27 were necessary for binding RNA. Furthermore, the linker's flexible orientation, which is controlled by the hydrogen bond between Asn11 NδH and Arg27 O, appears to be a structural basis for NCp8 existing as a multi-functional protein.

    Original languageEnglish
    Pages (from-to)269-277
    Number of pages9
    JournalJournal of biochemistry
    Volume141
    Issue number2
    DOIs
    Publication statusPublished - 2007 Feb

    Keywords

    • HIV
    • NMR
    • Nucleocapsid protein
    • RNA binding protein
    • Zinc finger

    ASJC Scopus subject areas

    • Biochemistry
    • Molecular Biology

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