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  • 1
    ISSN: 1573-5001
    Keywords: AIDS ; DYANA ; HIV-1 ; NMR structure ; ribonuclease H ; RNA-DNA hybrid ; torsion angle dynamics
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Abstract A high-quality NMR solution structure of the chimeric hybrid duplex r(gcaguggc)⋅r(gcca)d(CTGC) was determined using the program DYANA with its recently implemented new module FOUND, which performs exhaustive conformational grid searches for dinucleotides. To ensure conservative data interpretation, the use of 1H-1H lower distance limit constraints was avoided. The duplex comprises the tRNA–DNA junction formed during the initiation of HIV-1 reverse transcription. It forms an A-type double helix that exhibits distinct structural deviations from a standard A-conformation. In particular, the minor groove is remarkably narrow, and its width decreases from about 7.5 Å in the RNA/RNA stem to about 4.5 Å in the RNA/DNA segment. This is unexpected, since minor groove widths for A-RNA and RNA/DNA hybrid duplexes of ∼11 Å and ∼8.5 Å, respectively, were previously reported. The present, new structure supports that reverse transcriptase-associated RNaseH specificity is related primarily to conformational adaptability of the nucleic acid in 'induced-fit'-type interactions, rather than the minor groove width of a predominantly static nucleic acid duplex.
    Type of Medium: Electronic Resource
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  • 2
    ISSN: 1573-5001
    Keywords: Vicinal spin-spin coupling constants ; J-modulated [15N,1H]-COSY ; Nonlinear fit of J-modulation ; Protein conformation ; NMR structure of proteins
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Summary Improved experimental schemes for the recently introduced J-modulated [15N,1H]-correlation experiment for measurements of the homonuclear amide proton-Cα proton vicinal coupling constants.3JHNα, in uniformly15N-labeled proteins are described, and a nonlinear fit procedure is presented for quantitative evaluation of3JHNα. The method was first tested with the N-terminal DNA-binding domain of the 434 repressor (M=7.3 kDa), where at 13 C precise values of3JHNα in the range 2.0–9.5 Hz were obtained for all residues with resolved15N-1H cross peaks. It was then applied to theAntennapedia homeodomain complexed to a synthetic 14-base pair DNA fragment (molecular weight of the complex ∼ 18 kDa). The3JHNα values measured were found to be in excellent agreement with those predicted from the secondary structure of this protein in the complex.
    Type of Medium: Electronic Resource
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