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Crystal structure of Rac1 bound to its effector phospholipase C-β2

Nature Structural & Molecular Biology volume 13, pages 1135–1140 (2006)Cite this article

Abstract

Although diverse signaling cascades require the coordinated regulation of heterotrimeric G proteins and small GTPases, these connections remain poorly understood. We present the crystal structure of the GTPase Rac1 bound to phospholipase C-β2 (PLC-β2), a classic effector of heterotrimeric G proteins. Rac1 engages the pleckstrin-homology (PH) domain of PLC-β2 to optimize its orientation for substrate membranes. Gβγ also engages the PH domain to activate PLC-β2, and these two activation events are compatible, leading to additive stimulation of phospholipase activity. In contrast to PLC-δ, the PH domain of PLC-β2 cannot bind phosphoinositides, eliminating this mode of regulation. The structure of the Rac1–PLC-β2 complex reveals determinants that dictate selectivity of PLC-β isozymes for Rac GTPases over other Rho-family GTPases, and substitutions within PLC-β2 abrogate its stimulation by Rac1 but not by Gβγ, allowing for functional dissection of this integral signaling node.

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Figure 1: Structure of activated Rac1 in complex with PLC-β2.
Figure 2: Intermolecular interface of Rac1 and PLC-β2.
Figure 3: Electrostatic interactions restrict the specificity of GTPases for PLC-β2.
Figure 4: Mutagenesis of key interface residues reduces Rac-stimulated PLC-β2 activity.

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Acknowledgements

We thank S. Hicks for her valuable work on the apo structure of PLC-β2. This work was funded by the US National Institutes of Health (GM 057391).

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Authors and Affiliations

  1. Department of Biochemistry and Biophysics The University of North Carolina School of Medicine, Chapel Hill, 27599, North Carolina, USA

    Mark R Jezyk & John Sondek

  2. Department of Pharmacology The University of North Carolina School of Medicine, Chapel Hill, 27599, North Carolina, USA

    Jason T Snyder, Svetlana Gershberg, David K Worthylake, T Kendall Harden & John Sondek

  3. Lineberger Comprehensive Cancer Center, The University of North Carolina School of Medicine, Chapel Hill, 27599, North Carolina, USA

    David K Worthylake & John Sondek

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  1. Mark R Jezyk
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Contributions

M.R.J., T.K.H. and J.S. conceived, analyzed and performed experiments and cowrote the manuscript. J.T.S. and S.G. assisted with construct design. J.T.S. and D.K.W. assisted with X-ray data collection and phasing.

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Correspondence to John Sondek.

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The authors declare no competing financial interests.

Supplementary information

Supplementary Fig. 1

Schematic of PH domain regulation. (PDF 8150 kb)

Supplementary Fig. 2

Sequence alignments of Rho GTPases and phospholipase C isozymes. (PDF 227 kb)

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Jezyk, M., Snyder, J., Gershberg, S. et al. Crystal structure of Rac1 bound to its effector phospholipase C-β2. Nat Struct Mol Biol 13, 1135–1140 (2006). https://doi.org/10.1038/nsmb1175

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