Premium
Chemical Genetic Approaches for Studying DNA Replication
Author(s) -
McHenry Charles S,
Dallmann Garry
Publication year - 2008
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.22.1_supplement.264.2
Subject(s) - computational biology , dna , dna replication , biology , replication (statistics) , chemical biology , identification (biology) , chemistry , biochemistry , genetics , botany , virology
DNA replication of bacterial chromosomes is dependent upon approximately 20 essential proteins. We are taking a chemical biology approach to discover inhibitors of each stage of the replicative reaction. These inhibitors should be of general utility to the scientific community, both as specific blocks for replication reaction stages that can be used in cellular experiments and as specific mechanistic and structural probes for studies with purified proteins. The use of fully reconstituted reactions provides a particularly productive approach. Not only are the essential enzymatic activities targets, but also the various protein‐protein interactions and conformers associated with discrete kinetic steps. Inhibitor identification is accomplished by a global robotic screen for all targets in a single well of a microtiter plate coupled with a deconvolution strategy that guides us to the specific target. The use of reconstituted replicases from model Gram‐negative and Gram‐positive organisms permits immediate identification of compounds with broad spectrum potential. A battery of specificity assays permit weeding out compounds that act by non‐specific mechanisms.