Replication protein A (RPA) is a heterotrimeric, multifunctional protein that binds single-stranded DNA (ssDNA) and is essential for eukaryotic DNA metabolism. Using heteronuclear NMR methods we have investigated the domain interactions and ssDNA binding of a fragment from the 70 kDa subunit of human RPA (hRPA70). This fragment contains an N-terminal domain (NTD), which is important for hRPA70-protein interactions, connected to a ssDNA-binding domain (SSB1) by a flexible linker (hRPA701-3261). Correlation analysis of the amide 1H and 15 N chemical shifts was used to compare the structure of the NTD and SSB1 in hRPA701-326 with two smaller fragments that corresponded to the individual domains. High correlation coefficients verified that the NTD and SSB1 maintained their structures in hRPA701-326, indicating weak interdomain coupling. Weak interdomain coupling was also suggested by a comparison of the transverse relaxation rates for hRPA701-326 and one of the smaller hRPA70 fragments containing the NTD and the flexible linker (hRPA701-68).
Revised: March 3, 2005 |
Published: August 1, 2001
Citation
Daughdrill G.W., J. Ackerman, N.G. Isern, M.V. Botuyan, C.H. Arrowsmith, M.S. Wold, and D.F. Lowry. 2001.The Weak Interdomain Coupling Observed in the 70 kDa Subunit of Human Replication Protein A is Unaffected by ssDNA Binding.Nucleic Acids Research 29, no. 15:3270-3276.PNNL-SA-35978.