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Rac Regulates Its Effector Phospholipase Cγ(2) through Interaction with a Split Pleckstrin Homology Domain

Several isoforms of phospholipase C (PLC) are regulated through interactions with Ras superfamily GTPases, including Rac proteins. Interestingly, of two closely related PLCγ isoforms, only PLCγ(2) has previously been shown to be activated by Rac. Here, we explore the molecular basis of this interact...

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Autores principales: Walliser, Claudia, Retlich, Michael, Harris, Richard, Everett, Katy L., Josephs, Michelle B., Vatter, Petra, Esposito, Diego, Driscoll, Paul C., Katan, Matilda, Gierschik, Peter, Bunney, Tom D.
Formato: Texto
Lenguaje:English
Publicado: American Society for Biochemistry and Molecular Biology 2008
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2573054/
https://www.ncbi.nlm.nih.gov/pubmed/18728011
http://dx.doi.org/10.1074/jbc.M803316200
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author Walliser, Claudia
Retlich, Michael
Harris, Richard
Everett, Katy L.
Josephs, Michelle B.
Vatter, Petra
Esposito, Diego
Driscoll, Paul C.
Katan, Matilda
Gierschik, Peter
Bunney, Tom D.
author_facet Walliser, Claudia
Retlich, Michael
Harris, Richard
Everett, Katy L.
Josephs, Michelle B.
Vatter, Petra
Esposito, Diego
Driscoll, Paul C.
Katan, Matilda
Gierschik, Peter
Bunney, Tom D.
author_sort Walliser, Claudia
collection PubMed
description Several isoforms of phospholipase C (PLC) are regulated through interactions with Ras superfamily GTPases, including Rac proteins. Interestingly, of two closely related PLCγ isoforms, only PLCγ(2) has previously been shown to be activated by Rac. Here, we explore the molecular basis of this interaction as well as the structural properties of PLCγ(2) required for activation. Based on reconstitution experiments with isolated PLCγ variants and Rac2, we show that an unusual pleckstrin homology (PH) domain, designated as the split PH domain (spPH), is both necessary and sufficient to effect activation of PLCγ(2) by Rac2. We also demonstrate that Rac2 directly binds to PLCγ(2) as well as to the isolated spPH of this isoform. Furthermore, through the use of NMR spectroscopy and mutational analysis, we determine the structure of spPH, define the structural features of spPH required for Rac interaction, and identify critical amino acid residues at the interaction interface. We further discuss parallels and differences between PLCγ(1) and PLCγ(2) and the implications of our findings for their respective signaling roles.
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spelling pubmed-25730542008-11-05 Rac Regulates Its Effector Phospholipase Cγ(2) through Interaction with a Split Pleckstrin Homology Domain Walliser, Claudia Retlich, Michael Harris, Richard Everett, Katy L. Josephs, Michelle B. Vatter, Petra Esposito, Diego Driscoll, Paul C. Katan, Matilda Gierschik, Peter Bunney, Tom D. J Biol Chem Mechanisms of Signal Transduction Several isoforms of phospholipase C (PLC) are regulated through interactions with Ras superfamily GTPases, including Rac proteins. Interestingly, of two closely related PLCγ isoforms, only PLCγ(2) has previously been shown to be activated by Rac. Here, we explore the molecular basis of this interaction as well as the structural properties of PLCγ(2) required for activation. Based on reconstitution experiments with isolated PLCγ variants and Rac2, we show that an unusual pleckstrin homology (PH) domain, designated as the split PH domain (spPH), is both necessary and sufficient to effect activation of PLCγ(2) by Rac2. We also demonstrate that Rac2 directly binds to PLCγ(2) as well as to the isolated spPH of this isoform. Furthermore, through the use of NMR spectroscopy and mutational analysis, we determine the structure of spPH, define the structural features of spPH required for Rac interaction, and identify critical amino acid residues at the interaction interface. We further discuss parallels and differences between PLCγ(1) and PLCγ(2) and the implications of our findings for their respective signaling roles. American Society for Biochemistry and Molecular Biology 2008-10-31 /pmc/articles/PMC2573054/ /pubmed/18728011 http://dx.doi.org/10.1074/jbc.M803316200 Text en Copyright © 2008, The American Society for Biochemistry and Molecular Biology, Inc. Author's Choice Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/3.0/) applies to Author Choice Articles
spellingShingle Mechanisms of Signal Transduction
Walliser, Claudia
Retlich, Michael
Harris, Richard
Everett, Katy L.
Josephs, Michelle B.
Vatter, Petra
Esposito, Diego
Driscoll, Paul C.
Katan, Matilda
Gierschik, Peter
Bunney, Tom D.
Rac Regulates Its Effector Phospholipase Cγ(2) through Interaction with a Split Pleckstrin Homology Domain
title Rac Regulates Its Effector Phospholipase Cγ(2) through Interaction with a Split Pleckstrin Homology Domain
title_full Rac Regulates Its Effector Phospholipase Cγ(2) through Interaction with a Split Pleckstrin Homology Domain
title_fullStr Rac Regulates Its Effector Phospholipase Cγ(2) through Interaction with a Split Pleckstrin Homology Domain
title_full_unstemmed Rac Regulates Its Effector Phospholipase Cγ(2) through Interaction with a Split Pleckstrin Homology Domain
title_short Rac Regulates Its Effector Phospholipase Cγ(2) through Interaction with a Split Pleckstrin Homology Domain
title_sort rac regulates its effector phospholipase cγ(2) through interaction with a split pleckstrin homology domain
topic Mechanisms of Signal Transduction
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2573054/
https://www.ncbi.nlm.nih.gov/pubmed/18728011
http://dx.doi.org/10.1074/jbc.M803316200
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