CONTENT OF THE PROJECT
029 SSB binds to single-stranded DNA (ssDNA) stimulating viral DNA replication. This project is aimend to yield a three-dimensional structure of the ssDNA-029 SSB complex by NMR.
I-Determination of the three-dimensional structure in solution of 029 SSB.
The size of 029 SSB in solution makes it a suitable system to be analyz by NMR. We propose to determine the three-dimensional structure of the complex by isotopic labeling with either 15N or 13C (Folkers et al., 1994). II-Identificatiora of amino acids involved in binding to ssDNA.
Spin-labeled oligonucleotides (Folkers et al., 1993) and
lanthanide-chelate complexes (van Duynhoven et al., 1993) should allow to obtain information about those protein regions and residues involved in binding to single-stranded DNA.
III-Determirnation of the three-dimensional structure in solution of the ssDNA-029 SSB complex.
To determine the three-dimensional structure of the ssDNA-029 SSB complex, the protein will be labeled with 15N or 13C, leaving unlabeled the ligand (i.e., at natural isotopic abundance).