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The BARD1 C-Terminal Domain Structure and Interactions with Polyadenylation Factor CstF-50
[Image: see text] The BARD1 N-terminal RING domain binds BRCA1 while the BARD1 C-terminal ankyrin and tandem BRCT repeat domains bind CstF-50 to modulate mRNA processing and RNAP II stability in response to DNA damage. Here we characterize the BARD1 structural biochemistry responsible for CstF-50 bi...
Autores principales: | , , , , , |
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Formato: | Texto |
Lenguaje: | English |
Publicado: |
American Chemical Society
2008
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Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2654182/ https://www.ncbi.nlm.nih.gov/pubmed/18842000 http://dx.doi.org/10.1021/bi801115g |
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author | Edwards, Ross A. Lee, Megan S. Tsutakawa, Susan E. Williams, R. Scott Tainer, John A. Glover, J. N. Mark |
author_facet | Edwards, Ross A. Lee, Megan S. Tsutakawa, Susan E. Williams, R. Scott Tainer, John A. Glover, J. N. Mark |
author_sort | Edwards, Ross A. |
collection | PubMed |
description | [Image: see text] The BARD1 N-terminal RING domain binds BRCA1 while the BARD1 C-terminal ankyrin and tandem BRCT repeat domains bind CstF-50 to modulate mRNA processing and RNAP II stability in response to DNA damage. Here we characterize the BARD1 structural biochemistry responsible for CstF-50 binding. The crystal structure of the BARD1 BRCT domain uncovers a degenerate phosphopeptide binding pocket lacking the key arginine required for phosphopeptide interactions in other BRCT proteins. Small angle X-ray scattering together with limited proteolysis results indicates that ankyrin and BRCT domains are linked by a flexible tether and do not adopt a fixed orientation relative to one another. Protein pull-down experiments utilizing a series of purified BARD1 deletion mutants indicate that interactions between the CstF-50 WD-40 domain and BARD1 involve the ankyrin-BRCT linker but do not require ankyrin or BRCT domains. The structural plasticity imparted by the ANK-BRCT linker helps to explain the regulated assembly of different protein BARD1 complexes with distinct functions in DNA damage signaling including BARD1-dependent induction of apoptosis plus p53 stabilization and interactions. BARD1 architecture and plasticity imparted by the ANK-BRCT linker are suitable to allow the BARD1 C-terminus to act as a hub with multiple binding sites to integrate diverse DNA damage signals directly to RNA polymerase. |
format | Text |
id | pubmed-2654182 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2008 |
publisher | American Chemical Society |
record_format | MEDLINE/PubMed |
spelling | pubmed-26541822009-03-20 The BARD1 C-Terminal Domain Structure and Interactions with Polyadenylation Factor CstF-50 Edwards, Ross A. Lee, Megan S. Tsutakawa, Susan E. Williams, R. Scott Tainer, John A. Glover, J. N. Mark Biochemistry [Image: see text] The BARD1 N-terminal RING domain binds BRCA1 while the BARD1 C-terminal ankyrin and tandem BRCT repeat domains bind CstF-50 to modulate mRNA processing and RNAP II stability in response to DNA damage. Here we characterize the BARD1 structural biochemistry responsible for CstF-50 binding. The crystal structure of the BARD1 BRCT domain uncovers a degenerate phosphopeptide binding pocket lacking the key arginine required for phosphopeptide interactions in other BRCT proteins. Small angle X-ray scattering together with limited proteolysis results indicates that ankyrin and BRCT domains are linked by a flexible tether and do not adopt a fixed orientation relative to one another. Protein pull-down experiments utilizing a series of purified BARD1 deletion mutants indicate that interactions between the CstF-50 WD-40 domain and BARD1 involve the ankyrin-BRCT linker but do not require ankyrin or BRCT domains. The structural plasticity imparted by the ANK-BRCT linker helps to explain the regulated assembly of different protein BARD1 complexes with distinct functions in DNA damage signaling including BARD1-dependent induction of apoptosis plus p53 stabilization and interactions. BARD1 architecture and plasticity imparted by the ANK-BRCT linker are suitable to allow the BARD1 C-terminus to act as a hub with multiple binding sites to integrate diverse DNA damage signals directly to RNA polymerase. American Chemical Society 2008-10-09 2008-11-04 /pmc/articles/PMC2654182/ /pubmed/18842000 http://dx.doi.org/10.1021/bi801115g Text en Copyright © 2008 American Chemical Society http://pubs.acs.org This is an open-access article distributed under the ACS AuthorChoice Terms & Conditions. Any use of this article, must conform to the terms of that license which are available at http://pubs.acs.org. |
spellingShingle | Edwards, Ross A. Lee, Megan S. Tsutakawa, Susan E. Williams, R. Scott Tainer, John A. Glover, J. N. Mark The BARD1 C-Terminal Domain Structure and Interactions with Polyadenylation Factor CstF-50 |
title | The BARD1 C-Terminal Domain Structure and Interactions with Polyadenylation Factor CstF-50 |
title_full | The BARD1 C-Terminal Domain Structure and Interactions with Polyadenylation Factor CstF-50 |
title_fullStr | The BARD1 C-Terminal Domain Structure and Interactions with Polyadenylation Factor CstF-50 |
title_full_unstemmed | The BARD1 C-Terminal Domain Structure and Interactions with Polyadenylation Factor CstF-50 |
title_short | The BARD1 C-Terminal Domain Structure and Interactions with Polyadenylation Factor CstF-50 |
title_sort | bard1 c-terminal domain structure and interactions with polyadenylation factor cstf-50 |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2654182/ https://www.ncbi.nlm.nih.gov/pubmed/18842000 http://dx.doi.org/10.1021/bi801115g |
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