Article: Binding linkage in a telomere DNA-protein complex at the ends of Oxytricha nova chromosomes.
2005 Volume 350, Issue 5, Page(s) 938–952
Abstract: Alpha and beta protein subunits of the telomere end binding protein from Oxytricha nova (OnTEBP) combine with telomere single strand DNA to form a protective cap at the ends of chromosomes. We tested how protein-protein interactions seen in the co- ... ...
| Abstract | Alpha and beta protein subunits of the telomere end binding protein from Oxytricha nova (OnTEBP) combine with telomere single strand DNA to form a protective cap at the ends of chromosomes. We tested how protein-protein interactions seen in the co-crystal structure relate to DNA binding through use of fusion proteins engineered as different combinations of domains and subunits derived from OnTEBP. Joining alpha and beta resulted in a protein that bound single strand telomere DNA with high affinity (K(D-DNA)=1.4 nM). Another fusion protein, constructed without the C-terminal protein-protein interaction domain of alpha, bound DNA with 200-fold diminished affinity (K(D-DNA)=290 nM) even though the DNA-binding domains of alpha and beta were joined through a peptide linker. Adding back the alpha C-terminal domain as a separate protein restored high-affinity DNA binding. The binding behaviors of these fusion proteins and the native protein subunits are consistent with cooperative linkage between protein-association and DNA-binding equilibria. Linking DNA-protein stability to protein-protein contacts at a remote site may provide a trigger point for DNA-protein disassembly during telomere replication when the single strand telomere DNA must exchange between a very stable OnTEBP complex and telomerase. |
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| MeSH term(s) | Animals ; Chromosomes/metabolism ; DNA/metabolism ; Multiprotein Complexes ; Oxytricha/chemistry ; Oxytricha/genetics ; Protein Binding ; Protein Subunits ; Protozoan Proteins ; Recombinant Fusion Proteins ; Telomere/metabolism ; Telomere-Binding Proteins/metabolism | |||||
| Chemical Substances | Multiprotein Complexes ; Protein Subunits ; Protozoan Proteins ; Recombinant Fusion Proteins ; Telomere-Binding Proteins ; DNA (9007-49-2) | |||||
| Language | English | |||||
| Publishing date | 2005-07-29 | |||||
| Publishing country | England | |||||
| Document type | Journal Article ; Research Support, N.I.H., Extramural ; Research Support, U.S. Gov't, P.H.S. | |||||
| ZDB-ID | 80229-3 | |||||
| ISSN | 1089-8638 ; 0022-2836 | |||||
| ISSN (online) | 1089-8638 | |||||
| ISSN | 0022-2836 | |||||
| DOI | 10.1016/j.jmb.2005.05.040 | |||||
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| Database | MEDical Literature Analysis and Retrieval System OnLINE |
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