Identifying Critical Non-Catalytic Residues that Modulate Protein Kinase A Activity
BACKGROUND: Distal interactions between discrete elements of an enzyme are critical for communication and ultimately for regulation. However, identifying the components of such interactions has remained elusive due to the delicate nature of these contacts. Protein kinases are a prime example of an e...
Autores principales: | , , , , |
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Formato: | Texto |
Lenguaje: | English |
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Public Library of Science
2009
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Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2650257/ https://www.ncbi.nlm.nih.gov/pubmed/19270744 http://dx.doi.org/10.1371/journal.pone.0004746 |
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author | Kennedy, Eileen J. Yang, Jie Pillus, Lorraine Taylor, Susan S. Ghosh, Gourisankar |
author_facet | Kennedy, Eileen J. Yang, Jie Pillus, Lorraine Taylor, Susan S. Ghosh, Gourisankar |
author_sort | Kennedy, Eileen J. |
collection | PubMed |
description | BACKGROUND: Distal interactions between discrete elements of an enzyme are critical for communication and ultimately for regulation. However, identifying the components of such interactions has remained elusive due to the delicate nature of these contacts. Protein kinases are a prime example of an enzyme with multiple regulatory sites that are spatially separate, yet communicate extensively for tight regulation of activity. Kinase misregulation has been directly linked to a variety of cancers, underscoring the necessity for understanding intramolecular kinase regulation. METHODOLOGY/PRINCIPAL FINDINGS: A genetic screen was developed to identify suppressor mutations that restored catalytic activity in vivo from two kinase-dead Protein Kinase A mutants in S. cerevisiae. The residues defined by the suppressors provide new insights into kinase regulation. Many of the acquired mutations were distal to the nucleotide binding pocket, highlighting the relationship of spatially dispersed residues in regulation. CONCLUSIONS/SIGNIFICANCE: The suppressor residues provide new insights into kinase regulation, including allosteric effects on catalytic elements and altered protein-protein interactions. The suppressor mutations identified in this study also share overlap with mutations identified from an identical screen in the yeast PKA homolog Tpk2, demonstrating functional conservation for some residues. Some mutations were independently isolated several times at the same sites. These sites are in agreement with sites previously identified from multiple cancer data sets as areas where acquired somatic mutations led to cancer progression and drug resistance. This method provides a valuable tool for identifying residues involved in kinase activity and for studying kinase misregulation in disease states. |
format | Text |
id | pubmed-2650257 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2009 |
publisher | Public Library of Science |
record_format | MEDLINE/PubMed |
spelling | pubmed-26502572009-03-09 Identifying Critical Non-Catalytic Residues that Modulate Protein Kinase A Activity Kennedy, Eileen J. Yang, Jie Pillus, Lorraine Taylor, Susan S. Ghosh, Gourisankar PLoS One Research Article BACKGROUND: Distal interactions between discrete elements of an enzyme are critical for communication and ultimately for regulation. However, identifying the components of such interactions has remained elusive due to the delicate nature of these contacts. Protein kinases are a prime example of an enzyme with multiple regulatory sites that are spatially separate, yet communicate extensively for tight regulation of activity. Kinase misregulation has been directly linked to a variety of cancers, underscoring the necessity for understanding intramolecular kinase regulation. METHODOLOGY/PRINCIPAL FINDINGS: A genetic screen was developed to identify suppressor mutations that restored catalytic activity in vivo from two kinase-dead Protein Kinase A mutants in S. cerevisiae. The residues defined by the suppressors provide new insights into kinase regulation. Many of the acquired mutations were distal to the nucleotide binding pocket, highlighting the relationship of spatially dispersed residues in regulation. CONCLUSIONS/SIGNIFICANCE: The suppressor residues provide new insights into kinase regulation, including allosteric effects on catalytic elements and altered protein-protein interactions. The suppressor mutations identified in this study also share overlap with mutations identified from an identical screen in the yeast PKA homolog Tpk2, demonstrating functional conservation for some residues. Some mutations were independently isolated several times at the same sites. These sites are in agreement with sites previously identified from multiple cancer data sets as areas where acquired somatic mutations led to cancer progression and drug resistance. This method provides a valuable tool for identifying residues involved in kinase activity and for studying kinase misregulation in disease states. Public Library of Science 2009-03-09 /pmc/articles/PMC2650257/ /pubmed/19270744 http://dx.doi.org/10.1371/journal.pone.0004746 Text en Kennedy 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 Kennedy, Eileen J. Yang, Jie Pillus, Lorraine Taylor, Susan S. Ghosh, Gourisankar Identifying Critical Non-Catalytic Residues that Modulate Protein Kinase A Activity |
title | Identifying Critical Non-Catalytic Residues that Modulate Protein Kinase A Activity |
title_full | Identifying Critical Non-Catalytic Residues that Modulate Protein Kinase A Activity |
title_fullStr | Identifying Critical Non-Catalytic Residues that Modulate Protein Kinase A Activity |
title_full_unstemmed | Identifying Critical Non-Catalytic Residues that Modulate Protein Kinase A Activity |
title_short | Identifying Critical Non-Catalytic Residues that Modulate Protein Kinase A Activity |
title_sort | identifying critical non-catalytic residues that modulate protein kinase a activity |
topic | Research Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2650257/ https://www.ncbi.nlm.nih.gov/pubmed/19270744 http://dx.doi.org/10.1371/journal.pone.0004746 |
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