NMR analysis of the architecture and functional remodeling of a modular multidomain protein, RPA.
Modular proteins with multiple domains tethered by flexible linkers have variable global architectures. Using the eukaryotic ssDNA binding protein, Replication Protein A (RPA), we demonstrate that NMR spectroscopy is a powerful tool to characterize the remodeling of architecture in different functional states. The first direct evidence is obtained for the remodeling of RPA upon binding ssDNA, including an alteration in the availability of the RPA32N domain that may help explain its damage-dependent phosphorylation.
Brosey
Chris A
CA
Department of Biochemistry, and Center for Structural Biology, Vanderbilt University, Nashville, Tennessee 37232-8725, USA.
Chagot
Marie-Eve
ME
Ehrhardt
Mark
M
Pretto
Dalyir I
DI
Weiner
Brian E
BE
Chazin
Walter J
WJ
D004277:DNA, Single-Stranded / Q000378:metabolism; D004268:DNA-Binding Proteins / Q000737:chemistry / Q000378:metabolism; D009682:Magnetic Resonance Spectroscopy / Q000379:methods*; D010766:Phosphorylation; D011485:Protein Binding; D011487:Protein Conformation; D051716:Replication Protein A / Q000737:chemistry* / Q000378:metabolism
D004277:DNA, Single-Stranded; D004268:DNA-Binding Proteins; D051716:Replication Protein A