title: Functional dynamics in replication protein A DNA binding and protein recruitment domains.
abstract: Replication Protein A (RPA) is an essential scaffold for many DNA processing machines; its function relies on its modular architecture. Here, we report (15)N-nuclear magnetic resonance heteronuclear relaxation analysis to characterize the movements of single-stranded (ss) DNA binding and protein interaction modules in the RPA70 subunit. Our results provide direct evidence for coordination of the motion of the tandem RPA70AB ssDNA binding domains. Moreover, binding of ssDNA substrate is found to cause dramatic reorientation and full coupling of inter-domain motion. In contrast, the RPA70N protein interaction domain remains structurally and dynamically independent of RPA70AB regardless of binding of ssDNA. This autonomy of motion between the 70N and 70AB modules supports a model in which the two binding functions of RPA are mediated fully independently, but remain differentially coordinated depending on the length of their flexible tethers. A critical role for linkers between the globular domains in determining the functional dynamics of RPA is proposed.
authors.affiliation: Departments of Biochemistry and Chemistry, Center for Structural Biology, Vanderbilt University, Nashville, TN 37232-8725, USA.
authors.forename: Chris A
authors.initials: CA
authors.lastname: Brosey
authors.affiliation: Departments of Biochemistry and Chemistry, Center for Structural Biology, Vanderbilt University, Nashville, TN 37232-8725, USA.
authors.forename: Sarah E
authors.initials: SE
authors.lastname: Soss
authors.affiliation: Department of Biological Sciences, Vanderbilt University, Nashville, TN 37232, USA.
authors.forename: Sonja
authors.initials: S
authors.lastname: Brooks
authors.affiliation: Department of Chemistry, Georgia State University, Atlanta, GA 30302-4098, USA.
authors.forename: Chunli
authors.initials: C
authors.lastname: Yan
authors.affiliation: Department of Chemistry, Georgia State University, Atlanta, GA 30302-4098, USA.
authors.forename: Ivaylo
authors.initials: I
authors.lastname: Ivanov
authors.affiliation: Department of Physics, Indian Institute of Science Education and Research (IISER), Mohali, Sector 81 Manauli PO, SAS Nagar, Punjab 140306, India.
authors.forename: Kavita
authors.initials: K
authors.lastname: Dorai
authors.affiliation: Departments of Biochemistry and Chemistry, Center for Structural Biology, Vanderbilt University, Nashville, TN 37232-8725, USA; Department of Chemistry, Vanderbilt University, Nashville, TN 37232, USA. Electronic address: walter.chazin@vanderbilt.edu.
authors.forename: Walter J
authors.initials: WJ
authors.lastname: Chazin
mesh_terms: D004247:DNA / Q000378:metabolism*; D056324:Diffusion Tensor Imaging; D040461:Gene Components; D008958:Models, Molecular; D056004:Molecular Dynamics Simulation; D019906:Nuclear Magnetic Resonance, Biomolecular; D011485:Protein Binding; D011487:Protein Conformation; D017434:Protein Structure, Tertiary; D051716:Replication Protein A / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*
chemical_list: D051716:Replication Protein A; D004247:DNA