Structure-function analysis of drug resistent HIV-reverse transcriptase having new improved selection properties

• Research unit: EU RFP
• Project number: 97.0593

Key words (English)

AIDS; HW-1; NMR structure; isotope labelling of DNA and RNA; RNA-DNA hybrid

Alternative project number (English)

EU project number: BMH4-CT97-2641

Research programs (English)

EU-programme: 4. Frame Research Programme - 4.2 Agriculture and agroindustry

Short description (English)

See abstract

Further information (English)

Full name of research-institution/enterprise:
ETH Zürich
Institut für Molekularbiologie und Biophysik

Partners and International Organizations (English)

Coordinator: Max-Planck-Gesellschaft (D)

Abstract (English)

During the first phase of this project the contribution from my laboratory consisted in the determination of the three-dimensional structure of a hybrid duplex that corresponds to the tRNA-.DNA junction formed during initiation of HIV-1 reverse transcription. The experience gained with this project made it clear that similar results on more complex nucleic acid structures, increased precision of similar structures, or investigations of such nucleic acid moieties in their interactions with proteins could only be obteined with the use of isotope-labelling of the nucleic acids with 13C and 15N..During the second phase of the project we therefore commissioned the synthesis of an isotope-labelled hybrid duplex that would be of direct interest for further investigations of the initiation of HIV-1 reverse transcription. In parallel we started NMR-techniques developments for the envisaged studies with this labelled hybrid duplex, for which we used isotope-labelled DNA that was already in our laboratory. The methods developments were highly successful and resulted in three publications. With regard to future studies of hybrid duplexes relating to HIV-1 transciption initiation, the most exciting advance is represented by the discovery of novel scalar coupling constants across the hydrogen bonds in DNA and RNA structures. Direct application with the transcription initiation system had to be deferred to a follow-up project, since the synthesis of the labelled compound commissioned outside of our laboratory could not be concluded before the end of the project period.

References in databases (English)

Swiss Database: Euro-DB of the
State Secretariat for Education and Research
Hallwylstrasse 4
CH-3003 Berne, Switzerland
Tel. [phone]
Swiss Project-Number: 97.0593