Neutron scattering and nuclear magnetic resonance spectroscopy structural studies of protein-DNA complexes

摘要

This is the final report of a one-year, Laboratory-Directed Research and Development (LDRD) project at the Los Alamos National Laboratory (LANL). The project sought to employ advanced biophysical measurements to study the structure of nucleosomes and the structure of origins of DNA replication. The fundamental repeating unit of human chromosomes is the nucleosome, which contains about 200 base pairs of DNA and 9 histone proteins. Genome replication is strictly associated with the reversible acetylations of histones that unfold chromatin to allow access of factors to origins of DNA replications. The authors have studied two major structural problems: (1) the effects of histone acetylation on nucleosome structure, and (2) the structure of DNA origins of replication. They have recently completed preliminary X-ray scattering experiments at Stanford on positioned nucleosomes with defined DNA sequence and length, histone composition and level of acetylation. These experiments have shown that lengths of the DNA and acetylations of the histone H4 result in nucleosome structural changes. To understand internucleosomal interactions and the roles of histone H1 the authors have made preliminary x-ray scatter studies on native dinucleosomes that have demonstrated the feasibility of these experiments. The DNA sequence of the yeast replication origin has been synthesized for structure determination by multi-dimensional NMR spectroscopy.