Assembly and analysis of conical models for the HIV-1 core

Science

Volumn 283, Issue 5398, 1999, Pages 80-83

  Ganser, Barbie K ^a   Li, Su ^a   Klishko, Victor Y ^a   Finch, John T ^b   Sundquist, Wesley I ^a  

^a UNIVERSITY OF UTAH SCHOOL OF MEDICINE   (United States)

^b Med Res Cncl of Molecular Biology   (United Kingdom)

Author keywords

[No Author keywords available]

Indexed keywords

NUCLEOCAPSID PROTEIN;

ARTICLE; HUMAN IMMUNODEFICIENCY VIRUS 1; MOLECULAR DYNAMICS; NONHUMAN; PRIORITY JOURNAL; VIRUS ASSEMBLY; VIRUS CORE; VIRUS GENOME; VIRUS NUCLEOCAPSID;

CAPSID; GENE PRODUCTS, GAG; HIV-1; MATHEMATICS; MICROSCOPY, ELECTRON; MODELS, BIOLOGICAL; NUCLEOCAPSID; RECOMBINANT FUSION PROTEINS; RNA, VIRAL; TEMPLATES, GENETIC;

HUMAN IMMUNODEFICIENCY VIRUS; HUMAN IMMUNODEFICIENCY VIRUS 1; MIRIDAE;

EID: 0032930797     PISSN: 00368075     EISSN: None     Source Type: Journal    
DOI: 10.1126/science.283.5398.80     Document Type: Article

References (41)

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22. NL4-3 genome and the ribosomal RNA (control) was a 1441-nt fragment from 16S ribosomal RNA of B. stearothemophilus. The 6400-nt tobacco mosaic viral RNA was obtained by phenol and chloroform extractions of the virus and precipitated from ethanol CA-NC assembly reactions in the presence of noncognate RNAs were identical to those given in (9). In the absence of RNA, CA-NC cones formed under the following conditions: 300 μM CA-NC, 1 M NaCl, and 50 mM tris-HCl (pH 8.0) at 37°C for 60 min. In the absence of exogenous RNA, neither cones nor cylinders formed at concentrations of 0.5 M NaCl or below. Absorption spectra demonstrated that our CA-NC preparations were not contaminated with Escherichia coli RNA (estimated lower detection limit was ∼1 base/protein molecule). To control for even lower levels of RNA contamination, we preincubated the CA-NC protein with 0.5 mg/ml ribonuclease A (Type 1-AS, 54 Kunitz U/mg, Sigma) for 1 hour at 4°C, which then formed cones normally.
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40. Single-letter abbreviations for the amino acid residues are as follows: A, Ala; C, Cys; D, Asp; E, Glu; F, Phe; G, Gly; H, His; I, Ile; K, Lys; L, Leu; M, Met; N, Asn; P, Pro; Q, Gln; R, Arg; S, Ser; T, Thr; V, Val; W, Trp; and Y, Tyr.
41. We thank C. Hill for very helpful discussions on the relationship between viral cores and fullerene cones, D. Hobbs for refining the ChemDraw3D images of cones, G. Stubbs for a gift of tobacco mosiac virus, J. McCutcheon for the plasmid used to prepare ribosomal RNA, and K. Albertine and N. Chandler of the University of Utah Shared Electron Microscopy facility for their support and encouragement. Supported by grants from NIH and from the Huntsman Cancer Institute (to W.I.S.).