Secondary structure in solution of two anti-HIV-1 hammerhead ribozymes as investigated by two-dimensional1H 500 MHz NMR spectroscopy in water
- 9 January 1995
- journal article
- Published by Wiley in FEBS Letters
- Vol. 357 (3) , 317-323
- https://doi.org/10.1016/0014-5793(94)01388-h
Abstract
Two hammerhead chimeric RNA/DNA ribozymes (HRz) were synthesized in pure form. Both were 30 nucleotides long, and the sequences were such that they could be targeted to cleave the HIV-1 gag RNA. Named HRz-W and HRz-M, the former had its invariable core region conserved, the latter had a uridine in the invariable region replaced by a guanine. Their secondary structures were determined by 2D NOESY 1H 500 MHz NMR spectroscopy in 90% water and 10% D20, following the imino protons. The data show that both HRz-M and HRz-W form identical secondary structures with stem regions consisting of continuous stacks of AT and GT pairs. An energy mimimized computer model of this stem region is provided. The results suggest that the loss of catalytic activity that is known to result when an invariant core residue is replaced is not related to the secondary structure of the ribozymes in the absence of substrateKeywords
This publication has 15 references indexed in Scilit:
- A Three-Dimensional Model for the Hammerhead Ribozyme Based on Fluorescence MeasurementsScience, 1994
- Three-dimensional structure of a hammerhead ribozymeNature, 1994
- Chemical Synthesis, Analysis, and Purification of OligoribonucleotidesMethods, 1993
- Frontiers in HIV-1 Therapy: Fourth Conference of the NIAID National Cooperative Drug Discovery Groups-HIVAIDS Research and Human Retroviruses, 1992
- Systematic study of nuclear Overhauser effects vis-a-vis local helical parameters, sugar puckers, and glycosidic torsions in B DNA; insensitivity of NOE to local transitions in B DNA oligonucleotides due to internal structural compensationsBiochemistry, 1992
- MOLSCRIPT: a program to produce both detailed and schematic plots of protein structuresJournal of Applied Crystallography, 1991
- Sequence requirements of the hammerhead RNA self-cleavage reactionBiochemistry, 1990
- Ribozymes as Potential Anti-HIV-1 Therapeutic AgentsScience, 1990
- DNA structure in which an adenine-cytosine mismatch pair forms an integral part of the double helixBiochemistry, 1987
- Further refinement of the structure of yeast tRNAPheJournal of Molecular Biology, 1978