EUROFAN: European network for functional analysis of yeast genes discovered by systematic DNA sequencing

• Research unit: EU RFP
• Project number: 95.0191-11

Key words (English)

Chromatin structure; yeast; chromosomes; genes

Alternative project number (English)

EU project number: BIO4-CT95-0080

Research programs (English)

EU-programme: 4. Frame Research Programme - 4.1 Biotechnology

Short description (English)

See abstract

Partners and International Organizations (English)

P. Philippsen Basel

Abstract (English)

The 6-Pack Analysis (functional analysis of 6 genes) was completed as reported previously. This part of the project was done by Dr. S. TeHeesen (Prof. M. Aebi, Microbiology, ETH-Zürich).
In the chromatin project, it was proposed to study chromatin structure, in particular, the arrangement of nucleosomes and gene promoter regions along yeast chromosomes using footprinting techniques. The goals were: 1. To investigate whether techniques of nucleosome mapping may be developed or improved to allow mapping over long distances of several thousand base pairs. We did not find appropriate nuclease digestion and gel conditions to directly map long distances. However the membranes applied in th standard protocol can be reused several fold to map additional regions in the genome. 2. Evaluation of criterias for the interpretation, characterization and mapping. We discovered that the photoreactivating enzyme (DNA-photolyase), which repairs UV induced DNA-lesions, is tightly regulated by chromatin structure. This enzyme can now be used as an in vivo approach to measure structural and dynamic properties in chromosomes. While the first experiments were done in artificial minichromosomes, the protocol is now established to work in chromosomes. Since the results obtained by photolyase are fully compatible with the nuclease data, the quality of the chromatin preparation and the nuclease digestion protocols are highly reliable and can be used for future routine work.
Publication: Livingstone-Zatchej, M., Suter, B. and Thoma, F. (1999) Photolyase - a molecular tool to characterize chromatin structure in yeast. Methods in Molecular Biology 'Chromatin' (ed. P. Becker), in press.

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: 95.0191-11