       Document 0595
 DOCN  M9550595
 TI    The central globular domain of the nucleocapsid protein of human
       immunodeficiency virus type 1 is critical for virion structure and
       infectivity.
 DT    9505
 AU    Ottmann M; Gabus C; Darlix JL; LaboRetro, Unite de Virologie Humaine,
       Ecole Normale; Superieure-Institut National de la Sante et de la
       Recherche; Medicale, Lyon, France.
 SO    J Virol. 1995 Mar;69(3):1778-84. Unique Identifier : AIDSLINE
       MED/95156609
 AB    The nucleocapsid protein NCp7 of human immunodeficiency virus type 1
       (HIV-1) is a 72-amino-acid peptide containing two CCHC-type zinc fingers
       linked by a short basic sequence, 29RAPRKKG35, which is conserved in
       HIV-1 and simian immunodeficiency virus. The complete three-dimensional
       structure of NCp7 has been determined by 1H-nuclear magnetic resonance
       spectroscopy (N. Morellet, H. de Rocquigny, Y. Mely, N. Jullian, H.
       Demene, M. Ottmann, D. Gerard, J. L. Darlix, M. C. Fournie-Zaluski, and
       B. P. Roques, J. Mol. Biol. 235:287-301, 1994) and revealed a central
       globular domain where the two zinc fingers are brought in close
       proximity by the RAPRKKG linker. To examine the role of this globular
       structure and more precisely of the RAPRKKG linker in virion structure
       and infectivity, we generated HIV-1 DNA mutants in the RAPRKK sequence
       of NCp7 and analyzed the mutant virions produced by transfected cells.
       Mutations that probably alter the structure of NCp7 structure led to the
       formation of very poorly infectious virus (A30P) or noninfectious virus
       (P31L and R32G). In addition, the P31L mutant did not contain detectable
       amounts of reverse transcriptase and had an immature core morphology, as
       determined by electron microscopy. On the other hand, mutations changing
       the basic nature of NCp7 had poor effect. R29S had a wild-type
       phenotype, and the replacement of 32RKK34 by SSS (S3 mutant) resulted in
       a decrease by no more than 100-fold of the virus titer. These results
       clearly show that the RAPRKKG linker contains residues that are critical
       for virion structure and infectivity.
 DE    Amino Acid Sequence  Animal  Capsid/CHEMISTRY/*PHYSIOLOGY  Cell Line
       Cercopithecus aethiops  DNA Primers/CHEMISTRY  Gene Products,
       gag/CHEMISTRY/*PHYSIOLOGY  HIV-1/CHEMISTRY/PATHOGENICITY/*ULTRASTRUCTURE
       In Vitro  Molecular Sequence Data  Morphogenesis  Mutagenesis,
       Site-Directed  RNA, Viral/METABOLISM  Structure-Activity Relationship
       Support, Non-U.S. Gov't  Zinc Fingers  JOURNAL ARTICLE

       SOURCE: National Library of Medicine.  NOTICE: This material may be
       protected by Copyright Law (Title 17, U.S.Code).

