Determinants of Homodimerization Specificity in Histidine Kinases
Two-component signal transduction pathways consisting of a histidine kinase and a response regulator are used by prokaryotes to respond to diverse environmental and intracellular stimuli. Most species encode numerous paralogous histidine kinases that exhibit significant structural similarity. Yet in...
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| 语言: | en_US |
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Elsevier
2015
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| 在线阅读: | http://hdl.handle.net/1721.1/99134 https://orcid.org/0000-0002-8288-7607 https://orcid.org/0000-0003-4074-8980 |
| _version_ | 1826210792423292928 |
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| author | Ashenberg, Orr Rozen-Gagnon, Kathryn Keating, Amy E. Laub, Michael T |
| author2 | Massachusetts Institute of Technology. Computational and Systems Biology Program |
| author_facet | Massachusetts Institute of Technology. Computational and Systems Biology Program Ashenberg, Orr Rozen-Gagnon, Kathryn Keating, Amy E. Laub, Michael T |
| author_sort | Ashenberg, Orr |
| collection | MIT |
| description | Two-component signal transduction pathways consisting of a histidine kinase and a response regulator are used by prokaryotes to respond to diverse environmental and intracellular stimuli. Most species encode numerous paralogous histidine kinases that exhibit significant structural similarity. Yet in almost all known examples, histidine kinases are thought to function as homodimers. We investigated the molecular basis of dimerization specificity, focusing on the model histidine kinase EnvZ and RstB, its closest paralog in Escherichia coli. Direct binding studies showed that the cytoplasmic domains of these proteins each form specific homodimers in vitro. Using a series of chimeric proteins, we identified specificity determinants at the base of the four-helix bundle in the dimerization and histidine phosphotransfer domain. Guided by molecular coevolution predictions and EnvZ structural information, we identified sets of residues in this region that are sufficient to establish homospecificity. Mutating these residues in EnvZ to the corresponding residues in RstB produced a functional kinase that preferentially homodimerized over interacting with EnvZ. EnvZ and RstB likely diverged following gene duplication to yield two homodimers that cannot heterodimerize, and the mutants we identified represent possible evolutionary intermediates in this process. |
| first_indexed | 2024-09-23T14:55:46Z |
| format | Article |
| id | mit-1721.1/99134 |
| institution | Massachusetts Institute of Technology |
| language | en_US |
| last_indexed | 2024-09-23T14:55:46Z |
| publishDate | 2015 |
| publisher | Elsevier |
| record_format | dspace |
| spelling | mit-1721.1/991342022-09-29T11:27:44Z Determinants of Homodimerization Specificity in Histidine Kinases Ashenberg, Orr Rozen-Gagnon, Kathryn Keating, Amy E. Laub, Michael T Massachusetts Institute of Technology. Computational and Systems Biology Program Massachusetts Institute of Technology. Department of Biology Ashenberg, Orr Laub, Michael T. Keating, Amy E. Two-component signal transduction pathways consisting of a histidine kinase and a response regulator are used by prokaryotes to respond to diverse environmental and intracellular stimuli. Most species encode numerous paralogous histidine kinases that exhibit significant structural similarity. Yet in almost all known examples, histidine kinases are thought to function as homodimers. We investigated the molecular basis of dimerization specificity, focusing on the model histidine kinase EnvZ and RstB, its closest paralog in Escherichia coli. Direct binding studies showed that the cytoplasmic domains of these proteins each form specific homodimers in vitro. Using a series of chimeric proteins, we identified specificity determinants at the base of the four-helix bundle in the dimerization and histidine phosphotransfer domain. Guided by molecular coevolution predictions and EnvZ structural information, we identified sets of residues in this region that are sufficient to establish homospecificity. Mutating these residues in EnvZ to the corresponding residues in RstB produced a functional kinase that preferentially homodimerized over interacting with EnvZ. EnvZ and RstB likely diverged following gene duplication to yield two homodimers that cannot heterodimerize, and the mutants we identified represent possible evolutionary intermediates in this process. National Institutes of Health (U.S.) (Award GM067681) National Science Foundation (U.S.) (CAREER Grant) National Science Foundation (U.S.). Graduate Research Fellowship 2015-10-02T17:24:18Z 2015-10-02T17:24:18Z 2011-08 2011-08 Article http://purl.org/eprint/type/JournalArticle 00222836 1089-8638 http://hdl.handle.net/1721.1/99134 Ashenberg, Orr, Kathryn Rozen-Gagnon, Michael T. Laub, and Amy E. Keating. “Determinants of Homodimerization Specificity in Histidine Kinases.” Journal of Molecular Biology 413, no. 1 (October 2011): 222–235. https://orcid.org/0000-0002-8288-7607 https://orcid.org/0000-0003-4074-8980 en_US http://dx.doi.org/10.1016/j.jmb.2011.08.011 Journal of Molecular Biology Creative Commons Attribution-Noncommercial-NoDerivatives http://creativecommons.org/licenses/by-nc-nd/4.0/ application/pdf Elsevier PMC |
| spellingShingle | Ashenberg, Orr Rozen-Gagnon, Kathryn Keating, Amy E. Laub, Michael T Determinants of Homodimerization Specificity in Histidine Kinases |
| title | Determinants of Homodimerization Specificity in Histidine Kinases |
| title_full | Determinants of Homodimerization Specificity in Histidine Kinases |
| title_fullStr | Determinants of Homodimerization Specificity in Histidine Kinases |
| title_full_unstemmed | Determinants of Homodimerization Specificity in Histidine Kinases |
| title_short | Determinants of Homodimerization Specificity in Histidine Kinases |
| title_sort | determinants of homodimerization specificity in histidine kinases |
| url | http://hdl.handle.net/1721.1/99134 https://orcid.org/0000-0002-8288-7607 https://orcid.org/0000-0003-4074-8980 |
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