Solution structure of an “open” E. coli Pol III clamp loader sliding clamp complex

• Published in: Journal of structural biology 2016-06, Vol.194 (3), p.272-281
• Main Authors: Tondnevis, Farzaneh, Weiss, Thomas M., Matsui, Tsutomu, Bloom, Linda B., McKenna, Robert
• Format: Article
• Language: English
• Subjects: AAA+ ATPase; Adenosine Triphosphate - metabolism; Binding Sites; Chromatography, Gel; Clamp loader; DNA Polymerase III - metabolism; DNA Replication; DNA, Bacterial; DNA-Directed DNA Polymerase; Escherichia coli - enzymology; Escherichia coli - genetics; Escherichia coli Proteins; Models, Molecular; Protein Conformation; Protein Subunits; SAXS; Scattering, Small Angle; Sliding clamp; Solutions; X-Ray Diffraction
• ISSN: 1047-8477; 1095-8657
• DOI: 10.1016/j.jsb.2016.03.003

Sliding clamps are opened and loaded onto primer template junctions by clamp loaders, and once loaded on DNA, confer processivity to replicative polymerases. Previously determined crystal structures of eukaryotic and T4 clamp loader-clamp complexes have captured the sliding clamps in either closed or only partially open interface conformations. In these solution structure studies, we have captured for the first time the clamp loader-sliding clamp complex from Escherichia coli using size exclusion chromatography coupled to small angle X-ray scattering (SEC-SAXS). The data suggests the sliding clamp is in an open conformation which is wide enough to permit duplex DNA binding. The data also provides information about spatial arrangement of the sliding clamp with respect to the clamp loader subunits and is compared to complex crystal structures determined from other organisms.