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Distinct functional constraints driving conservation of the cofilin N-terminal regulatory tail
Cofilin family proteins have essential roles in remodeling the cytoskeleton through filamentous actin depolymerization and severing. The short unstructured N-terminal region of cofilin is critical for actin binding and harbors the major site of inhibitory phosphorylation. Atypically for a disordered...
Autores principales: | , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Cold Spring Harbor Laboratory
2023
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10327202/ https://www.ncbi.nlm.nih.gov/pubmed/37425676 http://dx.doi.org/10.1101/2023.06.30.547189 |
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author | Sexton, Joel A. Potchernikov, Tony Bibeau, Jeffrey P. Casanova-Sepúlveda, Gabriela Cao, Wenxiang Lou, Hua Jane Boggon, Titus J. De La Cruz, Enrique M. Turk, Benjamin E. |
author_facet | Sexton, Joel A. Potchernikov, Tony Bibeau, Jeffrey P. Casanova-Sepúlveda, Gabriela Cao, Wenxiang Lou, Hua Jane Boggon, Titus J. De La Cruz, Enrique M. Turk, Benjamin E. |
author_sort | Sexton, Joel A. |
collection | PubMed |
description | Cofilin family proteins have essential roles in remodeling the cytoskeleton through filamentous actin depolymerization and severing. The short unstructured N-terminal region of cofilin is critical for actin binding and harbors the major site of inhibitory phosphorylation. Atypically for a disordered sequence, the N-terminal region is highly conserved, but the aspects of cofilin functionality driving this conservation are not clear. Here, we screened a library of 16,000 human cofilin N-terminal sequence variants for their capacity to support growth in S. cerevisiae in the presence or absence of the upstream regulator LIM kinase. Results from the screen and subsequent biochemical analysis of individual variants revealed distinct sequence requirements for actin binding and regulation by LIM kinase. While the presence of a serine, rather than threonine, phosphoacceptor residue was essential for phosphorylation by LIM kinase, the native cofilin N-terminus was otherwise a suboptimal LIM kinase substrate. This circumstance was not due to sequence requirements for actin binding and severing, but rather appeared primarily to maintain the capacity for phosphorylation to inactivate cofilin. Overall, the individual sequence requirements for cofilin function and regulation were remarkably loose when examined separately, but collectively restricted the N-terminus to sequences found in natural cofilins. Our results illustrate how a regulatory phosphorylation site can balance potentially competing sequence requirements for function and regulation. |
format | Online Article Text |
id | pubmed-10327202 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2023 |
publisher | Cold Spring Harbor Laboratory |
record_format | MEDLINE/PubMed |
spelling | pubmed-103272022023-07-08 Distinct functional constraints driving conservation of the cofilin N-terminal regulatory tail Sexton, Joel A. Potchernikov, Tony Bibeau, Jeffrey P. Casanova-Sepúlveda, Gabriela Cao, Wenxiang Lou, Hua Jane Boggon, Titus J. De La Cruz, Enrique M. Turk, Benjamin E. bioRxiv Article Cofilin family proteins have essential roles in remodeling the cytoskeleton through filamentous actin depolymerization and severing. The short unstructured N-terminal region of cofilin is critical for actin binding and harbors the major site of inhibitory phosphorylation. Atypically for a disordered sequence, the N-terminal region is highly conserved, but the aspects of cofilin functionality driving this conservation are not clear. Here, we screened a library of 16,000 human cofilin N-terminal sequence variants for their capacity to support growth in S. cerevisiae in the presence or absence of the upstream regulator LIM kinase. Results from the screen and subsequent biochemical analysis of individual variants revealed distinct sequence requirements for actin binding and regulation by LIM kinase. While the presence of a serine, rather than threonine, phosphoacceptor residue was essential for phosphorylation by LIM kinase, the native cofilin N-terminus was otherwise a suboptimal LIM kinase substrate. This circumstance was not due to sequence requirements for actin binding and severing, but rather appeared primarily to maintain the capacity for phosphorylation to inactivate cofilin. Overall, the individual sequence requirements for cofilin function and regulation were remarkably loose when examined separately, but collectively restricted the N-terminus to sequences found in natural cofilins. Our results illustrate how a regulatory phosphorylation site can balance potentially competing sequence requirements for function and regulation. Cold Spring Harbor Laboratory 2023-07-09 /pmc/articles/PMC10327202/ /pubmed/37425676 http://dx.doi.org/10.1101/2023.06.30.547189 Text en https://creativecommons.org/licenses/by-nd/4.0/This work is licensed under a Creative Commons Attribution-NoDerivatives 4.0 International License (https://creativecommons.org/licenses/by-nd/4.0/) , which allows reusers to copy and distribute the material in any medium or format in unadapted form only, and only so long as attribution is given to the creator. The license allows for commercial use. |
spellingShingle | Article Sexton, Joel A. Potchernikov, Tony Bibeau, Jeffrey P. Casanova-Sepúlveda, Gabriela Cao, Wenxiang Lou, Hua Jane Boggon, Titus J. De La Cruz, Enrique M. Turk, Benjamin E. Distinct functional constraints driving conservation of the cofilin N-terminal regulatory tail |
title | Distinct functional constraints driving conservation of the cofilin N-terminal regulatory tail |
title_full | Distinct functional constraints driving conservation of the cofilin N-terminal regulatory tail |
title_fullStr | Distinct functional constraints driving conservation of the cofilin N-terminal regulatory tail |
title_full_unstemmed | Distinct functional constraints driving conservation of the cofilin N-terminal regulatory tail |
title_short | Distinct functional constraints driving conservation of the cofilin N-terminal regulatory tail |
title_sort | distinct functional constraints driving conservation of the cofilin n-terminal regulatory tail |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10327202/ https://www.ncbi.nlm.nih.gov/pubmed/37425676 http://dx.doi.org/10.1101/2023.06.30.547189 |
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