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Dynein and Dynactin Leverage Their Bivalent Character to Form a High-Affinity Interaction
Cytoplasmic dynein and dynactin participate in retrograde transport of organelles, checkpoint signaling and cell division. The principal subunits that mediate this interaction are the dynein intermediate chain (IC) and the dynactin p150(Glued); however, the interface and mechanism that regulates thi...
Autores principales: | , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Public Library of Science
2013
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3618186/ https://www.ncbi.nlm.nih.gov/pubmed/23577064 http://dx.doi.org/10.1371/journal.pone.0059453 |
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author | Siglin, Amanda E. Sun, Shangjin Moore, Jeffrey K. Tan, Sarah Poenie, Martin Lear, James D. Polenova, Tatyana Cooper, John A. Williams, John C. |
author_facet | Siglin, Amanda E. Sun, Shangjin Moore, Jeffrey K. Tan, Sarah Poenie, Martin Lear, James D. Polenova, Tatyana Cooper, John A. Williams, John C. |
author_sort | Siglin, Amanda E. |
collection | PubMed |
description | Cytoplasmic dynein and dynactin participate in retrograde transport of organelles, checkpoint signaling and cell division. The principal subunits that mediate this interaction are the dynein intermediate chain (IC) and the dynactin p150(Glued); however, the interface and mechanism that regulates this interaction remains poorly defined. Herein, we use multiple methods to show the N-terminus of mammalian dynein IC, residues 10–44, is sufficient for binding p150(Glued). Consistent with this mapping, monoclonal antibodies that antagonize the dynein-dynactin interaction also bind to this region of the IC. Furthermore, double and triple alanine point mutations spanning residues 6 to 19 in the yeast IC homolog, Pac11, produce significant defects in spindle positioning. Using the same methods we show residues 381 to 530 of p150(Glued) form a minimal fragment that binds to the dynein IC. Sedimentation equilibrium experiments indicate that these individual fragments are predominantly monomeric, but admixtures of the IC and p150(Glued) fragments produce a 2:2 complex. This tetrameric complex is sensitive to salt, temperature and pH, suggesting that the binding is dominated by electrostatic interactions. Finally, circular dichroism (CD) experiments indicate that the N-terminus of the IC is disordered and becomes ordered upon binding p150(Glued). Taken together, the data indicate that the dynein-dynactin interaction proceeds through a disorder-to-order transition, leveraging its bivalent-bivalent character to form a high affinity, but readily reversible interaction. |
format | Online Article Text |
id | pubmed-3618186 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2013 |
publisher | Public Library of Science |
record_format | MEDLINE/PubMed |
spelling | pubmed-36181862013-04-10 Dynein and Dynactin Leverage Their Bivalent Character to Form a High-Affinity Interaction Siglin, Amanda E. Sun, Shangjin Moore, Jeffrey K. Tan, Sarah Poenie, Martin Lear, James D. Polenova, Tatyana Cooper, John A. Williams, John C. PLoS One Research Article Cytoplasmic dynein and dynactin participate in retrograde transport of organelles, checkpoint signaling and cell division. The principal subunits that mediate this interaction are the dynein intermediate chain (IC) and the dynactin p150(Glued); however, the interface and mechanism that regulates this interaction remains poorly defined. Herein, we use multiple methods to show the N-terminus of mammalian dynein IC, residues 10–44, is sufficient for binding p150(Glued). Consistent with this mapping, monoclonal antibodies that antagonize the dynein-dynactin interaction also bind to this region of the IC. Furthermore, double and triple alanine point mutations spanning residues 6 to 19 in the yeast IC homolog, Pac11, produce significant defects in spindle positioning. Using the same methods we show residues 381 to 530 of p150(Glued) form a minimal fragment that binds to the dynein IC. Sedimentation equilibrium experiments indicate that these individual fragments are predominantly monomeric, but admixtures of the IC and p150(Glued) fragments produce a 2:2 complex. This tetrameric complex is sensitive to salt, temperature and pH, suggesting that the binding is dominated by electrostatic interactions. Finally, circular dichroism (CD) experiments indicate that the N-terminus of the IC is disordered and becomes ordered upon binding p150(Glued). Taken together, the data indicate that the dynein-dynactin interaction proceeds through a disorder-to-order transition, leveraging its bivalent-bivalent character to form a high affinity, but readily reversible interaction. Public Library of Science 2013-04-05 /pmc/articles/PMC3618186/ /pubmed/23577064 http://dx.doi.org/10.1371/journal.pone.0059453 Text en © 2013 Siglin et al http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited. |
spellingShingle | Research Article Siglin, Amanda E. Sun, Shangjin Moore, Jeffrey K. Tan, Sarah Poenie, Martin Lear, James D. Polenova, Tatyana Cooper, John A. Williams, John C. Dynein and Dynactin Leverage Their Bivalent Character to Form a High-Affinity Interaction |
title | Dynein and Dynactin Leverage Their Bivalent Character to Form a High-Affinity Interaction |
title_full | Dynein and Dynactin Leverage Their Bivalent Character to Form a High-Affinity Interaction |
title_fullStr | Dynein and Dynactin Leverage Their Bivalent Character to Form a High-Affinity Interaction |
title_full_unstemmed | Dynein and Dynactin Leverage Their Bivalent Character to Form a High-Affinity Interaction |
title_short | Dynein and Dynactin Leverage Their Bivalent Character to Form a High-Affinity Interaction |
title_sort | dynein and dynactin leverage their bivalent character to form a high-affinity interaction |
topic | Research Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3618186/ https://www.ncbi.nlm.nih.gov/pubmed/23577064 http://dx.doi.org/10.1371/journal.pone.0059453 |
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