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Scaffold Proteins: Hubs for Controlling the Flow of Cellular Information Matthew C Scaffold Proteins: Hubs for Controlling the Flow of Cellular Information Matthew C. Good, et al. Science 332, 680 (2011); DOI: 10.1126/science.1198701 This copy is for your personal, non-commercial use only. If you wish to distribute this article to others, you can order high-quality copies for your colleagues, clients, or customers by clicking here. Permission to republish or repurpose articles or portions of articles can be obtained by following the guidelines here. The following resources related to this article are available online at www.sciencemag.org (this infomation is current as of May 25, 2011 ): Updated information and services, including high-resolution figures, can be found in the online version of this article at: http://www.sciencemag.org/content/332/6030/680.full.html Supporting Online Material can be found at: http://www.sciencemag.org/content/suppl/2011/05/04/332.6030.680.DC1.html on May 25, 2011 This article cites 57 articles, 28 of which can be accessed free: http://www.sciencemag.org/content/332/6030/680.full.html#ref-list-1 This article appears in the following subject collections: Cell Biology http://www.sciencemag.org/cgi/collection/cell_biol www.sciencemag.org Downloaded from Science (print ISSN 0036-8075; online ISSN 1095-9203) is published weekly, except the last week in December, by the American Association for the Advancement of Science, 1200 New York Avenue NW, Washington, DC 20005. Copyright 2011 by the American Association for the Advancement of Science; all rights reserved. The title Science is a registered trademark of AAAS. REVIEW used in evolution or engineering to change sig- naling behaviors. Scaffold Proteins: Versatile Tools to Scaffold Proteins: Hubs for Controlling Assemble Diverse Pathways Scaffolds are extremely diverse proteins, many of the Flow of Cellular Information which are likely to have evolved independently. Nonetheless they are conceptually related, in that Matthew C. Good,1* Jesse G. Zalatan,1 Wendell A. Lim1,2,3† they are usually composed of multiple modular interaction domains or motifs (see Box 1A for an example). Their exact domain composition and The spatial and temporal organization of molecules within a cell is critical for coordinating order, however, can vary widely depending on the many distinct activities carried out by the cell. In an increasing number of biological the pathways that they organize. signaling processes, scaffold proteins have been found to play a central role in physically In some cases, homologous individual inter- assembling the relevant molecular components. Although most scaffolds use a simple tethering action motifs can be found in scaffolds associated mechanism to increase the efficiency of interaction between individual partner molecules, with particular signaling proteins. For example, these proteins can also exert complex allosteric control over their partners and are themselves the AKAPs (A-kinase anchoring proteins), which the target of regulation. Scaffold proteins offer a simple, flexible strategy for regulating selectivity in pathways, shaping output behaviors, and achieving new responses from preexisting 1Department of Cellular and Molecular Pharmacology, Uni- signaling components. As a result, scaffold proteins have been exploited by evolution, versity of California, San Francisco, CA 94158, USA. 2Center for pathogens, and cellular engineers to reshape cellular behavior. Systems and Synthetic Biology, University of California, San Francisco, CA 94158, USA. 3Howard Hughes Medical Institute, University of California, San Francisco, CA 94158, USA. ammalian cells contain an estimated of pathways that they coordinate, the ways that *Present address: Department of Molecular and Cellular Bi- ology, University of California, Berkeley, CA 94720, USA. 1billionindividualproteinmolecules, they shape signaling responses, their biochemical †To whom correspondence should be addressed. E-mail: on May 25, 2011 M with as many as 10% of these involved and structural mechanisms, and how they are [email protected] in signal transduction (1). Given this enormous number of mole- cules, it seems remarkable that cells A “Well-mixed” cell B Intracellular signaling scaffolds can accurately process the vast ar- Spatial organization ray of signaling information they Pheromone INPUT EGF constantly receive. How can sig- Compartmentalization naling proteins find their correct (organelle) GPCR Receptor partners—and avoid their incorrect ␥ partners—among so many other ␣ ␤ Ste20 Ras www.sciencemag.org proteins? Colocalization Ste11 MAPKKK Raf Aprinciplethathasemerged Ste5 KSR over the past two decades is that Scaffold-mediated Yeast Ste7 MAPKK MEK MammalianMamma cells achieve specificity in their complex assembly mating ERK response MAPK Erk pathway molecular signaling networks by Fus3 organizing discrete subsets of pro- C teins in space and time (Fig. 1A) Cell-cell signaling junctions Mating Proliferation Downloaded from (2–4). For example, functionally OUTPUT interacting signaling components can be sequestered into specific AMPA regulatory NMDA proteins (TARPs, subcellular compartments (e.g., receptors AMPA organelles) or at the plasma mem- e.g. stargazin) receptors D Assembly-line processes brane. Another solution is to assem- ble functionally interacting proteins Unfolded into specific complexes. More than protein PDZ PDZ PDZ SH3-GK PSD-95 HOP 15 years ago, the first scaffold pro- NeuronalNeuronal sysynapse Hsp70 Sequentialquential teins were discovered—proteins chaperone that coordinate the physical as- Other reactions Hsp90 Folded sembly of components of a sig- Downstream scaffolds protein naling pathway or network (5–10). signaling effectors Actin cytoskeleton These proteins have captured the (e.g. nNOS) attention of the signaling field be- Fig. 1. Scaffold proteins organize cellular information flow. (A)Spatialorganizationisnecessary to achieve high-fidelity cause they appear to provide a intracellular information transfer. Proteins can be assembled into specificcomplexesbycompartmentalization simple and elegant solution for (organelle targeting), by membrane localization, and by scaffold proteins. (B)Intracellularsignalingpathwaysoftenuse determining the specificity of in- scaffold proteins. Canonical examples include Ste5, essential to the yeast mating MAPK pathway, and KSR, which formation flow in intracellular directs signaling in the mammalian Ras-Raf-MEK-MAPK pathway. (C)Scaffoldproteinsalsoplayanimportantrolein networks (2, 11, 12). We review organizing cell-cell communication junctions, such as neuronal synapses.ThePDZscaffold,PSD-95,controlsNMDAand our current understanding of these AMPA glutamate receptor targeting to the synapse. (D)Assembly-lineprocessessuchas protein folding use scaffold organizational proteins: the types proteins. The HOP protein promotes transfer of unfolded proteins between Hsp70 and Hsp90 chaperones. 680 6 MAY 2011 VOL 332 SCIENCE www.sciencemag.org REVIEW A Modular interactions of Ste5 scaffold protein Three-tiered kinase cascade Gβ (Ste4) Fus3 PIP2 Ste11 Ste7 Fus3 Plasma Heterotrimeric MAPK Lipid MAPKKK MAPKK MAPK membrane G protein (regulatory) (catalytic) LIGANDS Ste5 + + + + + + + + (917 aa) N C - - - - - - - - - BINDING (463-514) DOMAINS Ste11-BD PM RING Fus3-BD PH Ste5ms (VWA domain) (45-64) (177-229) (288-315) (388-518) (593-786) Input Core signaling regulated Allosteric pathway Membrane recruitment Tether MAPK cascade coactivation of Fus3 Higher-order Target of Cdk Mediates feedback FUNCTION regulation negative regulation phosph. of Ste5 by Fus3 on May 25, 2011 B Core steps of signal propagation C Higher-order regulatory mechanisms Feedback 2 Membrane Cdk negative regulation Morphological 1 Receptor recruitment 4 MAPK of pathway activation phosphorylation response (% of cells schmooing) activation Tether MAPK activation of Ste5 α-factor 3 100 cascade Ste5 INPUT Phosph. Gβγ Ste20 Gβγ Ste20 80 wild-type blocks 60 α βγ βγ βγ binding P Ste11 40 G G Ste20 G Ste20 G Ste20 P 20 www.sciencemag.org P P P Ste11 P Ste11 P P Ste7 0 P 100 P P Cdk on Fus3 Ste5 Ste11 Ste7 Ste7 Ste11 Fus3 80 ∆Fus3-BD 60 G1 G2 Ste7 Fus3 Ste7 40 P M 20 Fus3 Cdk off Fus3 Fus3 Fus3 0 OUTPUT α -Factor (µM) Downloaded from Box 1. Structure and mechanisms of a canonical scaffold: the MAPK 3FZE), and can be efficiently phosphorylated by the colocalized and scaffold protein Ste5. (A) The Ste5 scaffold protein is composed of activated MAPKKK Ste11. The minimal VWA also functions as a coac- modular interaction domains, some of which mediate essential steps in tivator that permits Ste7 activation of the MAPK Fus3, which is tethered the three-tiered mating MAPK signaling cascade, and some of which to Ste7 via docking motifs (PDB ID 2B9H) (18, 57) (see Fig. 4C). (C) function in higher-order regulatory behaviors. (B)Corestepsofmatingpath- The Ste5 scaffold also takes part in higher-order regulatory processes. way (see also movie S1): Binding of a-factor peptide to its receptor (Ste2) Phosphorylation of the PM helix by Cdk blocks Ste5 membrane binding leads to activation of the guanine nucleotide binding protein (G protein) (25), thus preventing activation of the mating response at specific stages and dissociation of Gbg (Ste4 and Ste18) from the Ga subunit (Gpa1). of the cell cycle. The Fus3-binding domain (Fus3-BD; PDB ID 2F49) The Ste5 RING domain binds to the free Gbg complex (54, 55), triggering appears to be important for regulatory feedback phosphorylation of rapid recruitment of the scaffold to the membrane. Stabilization and Ste5 by Fus3, rather
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