Document 0334
 DOCN  M9460334
 TI    Correlation of nucleotide base and sugar protons in a 15N-labeled HIV-1
       RNA oligonucleotide by 1H-15N HSQC experiments.
 DT    9404
 AU    Sklenar V; Peterson RD; Rejante MR; Feigon J; Department of Chemistry
       and Biochemistry, University of; California, Los Angeles 90024.
 SO    J Biomol NMR. 1994 Jan;4(1):117-22. Unique Identifier : AIDSLINE
       MED/94176987
 AB    The advent of methods for preparing 15N- and 13C-labeled RNA
       oligonucleotides holds promise for extending the size of RNA molecules
       that can be studied by NMR spectroscopy. A practical limitation is the
       expense of the 13C label. It may therefore sometimes be desirable to
       prepare a relatively inexpensive 15N-labeled sample only. Here we show
       that the two-bond 1H-15N HSQC experiment can be used on 15N-labeled RNA
       to correlate the intranucleotide H1' and H8,H6,H5 resonances indirectly
       through the shared glycosidic nitrogen. The nonrefocused version of a
       standard HSQC experiment for 2D proton-detected 1H-15N chemical-shift
       correlation is applied in order to minimize the sensitivity loss due to
       the relatively fast spin-spin relaxation of RN oligonucleotides. The
       experiment is applied to the 30-nucleotide RNA RBE3 which contains the
       high-affinity binding site of the RRE (rev response element) for the Rev
       protein of HIV. The results indicate that this simple experiment allows
       a straightforward identification of the base proton resonances CH5, CH6,
       UH5, UH6, purine H8, and AH2 as well as the intranucleotide H1' and
       H8,H6,H5 connectivities. When combined with a NOESY experiment, complete
       sequential assignments can be obtained.
 DE    Base Sequence  Carbohydrates/CHEMISTRY  Gene Products, rev/METABOLISM
       Glycosides/CHEMISTRY  Hydrogen/CHEMISTRY  HIV-1/*CHEMISTRY/GENETICS
       Molecular Sequence Data  Nitrogen/CHEMISTRY  Nitrogen Isotopes  Nuclear
       Magnetic Resonance/*METHODS  Nucleotides/*CHEMISTRY
       Oligoribonucleotides/*CHEMISTRY  RNA, Viral/*CHEMISTRY/METABOLISM
       Support, U.S. Gov't, Non-P.H.S.  Support, U.S. Gov't, P.H.S.  JOURNAL
       ARTICLE

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