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Functional Plasticity of the BNIP-2 and Cdc42gap Homology

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Functional Plasticity of the BNIP-2 and Cdc42gap Homology FEBS Letters 586 (2012) 2674–2691 journal homepage: www.FEBSLetters.org Review Functional plasticity of the BNIP-2 and Cdc42GAP Homology (BCH) domain in cell signaling and cell dynamics ⇑ Catherine Qiurong Pan b, Boon Chuan Low a,b, a Cell Signaling and Developmental Biology Laboratory, Department of Biological Sciences, 14 Science Drive 4, National University of Singapore, 117543 Singapore, Singapore b Mechanobiology Institute Singapore, 5A Engineering Drive 1, National University of Singapore, 117411 Singapore, Singapore article info abstract Article history: The BNIP-2 and Cdc42GAP Homology (BCH) domains constitute a new and expanding family of Received 13 March 2012 highly conserved scaffold protein domains that regulate Rho, Ras and MAPK signaling, leading to Revised 16 April 2012 cell growth, apoptosis, morphogenesis, migration and differentiation. Such versatility is achieved Accepted 16 April 2012 via their ability to target small GTPases and their immediate regulators such as GTPase-activating Available online 21 April 2012 proteins (GAPs) and guanine nucleotide exchange factors (GEFs), their ability to form intra- Edited by Marius Sudol, Gianni Cesareni, molecular or inter-molecular interaction with itself or with other BCH domains, and also by their Giulio Superti-Furga and Wilhelm Just ability to bind diverse cellular proteins such as membrane receptors, isomerase, caspases and metabolic enzymes such as glutaminase. The presence of BCH and BCH-like domains in various Keywords: proteins and their divergence from the ancestral lipid-binding CRAL–TRIO domain warrant the need BCH domain to examine closely their structural, functional and regulatory plasticity in isolation or in concert CRAL–TRIO domain with other protein modules present in the same proteins. GTPase Ó 2012 Federation of European Biochemical Societies. Published by Elsevier B.V. All rights reserved. Kinase Modular Scaffold Signaling Protein–protein interaction Dynamics 1. Introduction combination, and how their potential functions are regulated across molecular, cellular and tissue levels. For example, the 1.1. Modular protein domains defines specificity, integration and adaptor protein, Grb2 contains both Src-homology-2 (SH2) and crosstalk of signaling networks Src-homology-3 (SH3) domains which bridge interaction between the phosphorylated tyrosine residue on the receptor tyrosine Protein domains are the autonomous, minimal modular entities kinase and the core PXXP motif of the RasGEF, SOS, respectively, that constitute the basic structural or/and functional units of a thereby transducing signals upon binding of ligands at the extra- polypeptide. They can act as protein docking sites, enzymatic units cellular space to intracellular milieu [2]. Precise molecular recogni- or regulatory devices and are classified into different families tion is also important for transcriptional regulation. For example, according to their conserved primary sequences. Each family of do- under the HIPPO signaling which is crucial for the control of organ main recognizes unique interaction motif at their target sites. As size and tumor suppression, the LATS kinase via its PPxY motif, such, proteins exploit different architecture designs of modular do- targets the WW domain of the transcriptional regulator, Yes mains in order to exert common, overlapping or distinct functions, associated protein (YAP) to phosphorylate YAP, leading to seques- particularly in mediating protein–protein and protein–lipid inter- tration of YAP in cytoplasm by protein 14-3-3. This mechanism actions that form the basis for intracellular and intercellular signal- helps reduce the YAP-induced cell proliferation and tumor ing [1]. suppression [3]. Similarly, in protein trafficking and targeting, As cells sense both biochemical and physical environments, it is interaction between the pleckstrin homology (PH) domain of the important to define how these domains work in isolation and in molecular motor, dynamin and phosphoinositide is essential for endocytosis [4,5]. To further enhance the efficiency of multi-component signaling ⇑ Corresponding author at: Cell Signaling and Developmental Biology Laboratory, Department of Biological Sciences, 14 Science Drive 4, National University of networks, cells employ specific scaffold proteins that utilize their Singapore, 117543 Singapore, Singapore. Fax: +65 6872 6123. multiple protein domains to recruit and organize different targets E-mail address: [email protected] (B.C. Low). into higher order macromolecular complexes at specific location 0014-5793/$36.00 Ó 2012 Federation of European Biochemical Societies. Published by Elsevier B.V. All rights reserved. http://dx.doi.org/10.1016/j.febslet.2012.04.023 C.Q. Pan, B.C. Low / FEBS Letters 586 (2012) 2674–2691 2675 and to fine-tune the activity and crosstalk among them. For exam- tion often lead to developmental defects and disease, less is known ples, the activity of Raf-Mek-Erk or Mek-Erk nodes downstream of about how this ‘‘3G’’ (GTPase, GEF, GAP)-signalome itself is regu- Ras is supported by different scaffold proteins at different locales, lated by cellular factors and how their functions and regulation namely the KSR (cell membrane), MP1 (late endosome), Sef (Golgi), are linked to other signaling networks downstream of membrane paxillin (focal adhesions), IQGAP1 (cytoskeleton) and b-arrestin receptor and MAP kinase modules. We therefore hypothesized that (early endosome) [6]. Such distribution of signaling nodes by BCH domain in BNIP-2 and Cdc42GAP could serve as a binding distinct hubs ensure localized and more specific control in the acti- platform for GTPases and possibly their immediate regulators, or vation (and deactivation) of Erk during different cellular processes. they could form multimeric form of BNIP-2, as sometimes seen Last but not least, coordination of cell adhesion dynamics require for other homologous domains used in protein dimerization/ the FERM (four-point-one, ezrin, radizin, moesin) domain of oligomerization. FAK (focal adhesion kinase) interacting with phospholipids Based on structural modeling, site-directed mutagenesis, PtdIns(4,5)P2 at the cell membrane to release the autoinhibition binding studies and GTPase assays, we revealed that BCH do- in FAK. This facilitates the recruitment of Src tyrosine kinase and main of BNIP-2 indeed binds to itself and the homologous BCH formation of focal adhesion complex via integration of many more domain of Cdc42GAP via a unique motif, 217-RRKMP-221 [11]. scaffold proteins, kinases, phosphatases, GTPases and their Interestingly, BNIP-2 BCH domain also binds Cdc42, a partner immediate regulators [7,8]. of the Cdc42GAP, via a sequence motif similar to the CRIB Therefore, precise locales and timing for the molecular recogni- (Cdc42/Rac1-Interactive Binding) motif commonly present in tion by different combinations of protein domains in cis and in the effector proteins of Cdc42/Rac1 [16,17] (see Fig. 3A and C trans will help define the specificity, connectivity, crosstalk and for details later). Unlike binding of CRIB motif which is abso- feedback among various signaling nodes. Acting in concert, they lutely GTP-dependent (in order to allow activation of effectors regulate important functions such as cytoskeleton rearrangement, pathway), the BCH domain of BNIP-2 does not distinguish be- cell–cell adhesion, cell–matrix interaction, protein trafficking, tween the unloaded, GDP and GTP-bound form of Cdc42 organelle and membrane dynamics, cell metabolism and immune in vitro [9]. However, it binds preferentially to the dominant response, gene expression and protein synthesis and stability, lead- negative form of Cdc42, T17N but not with the constitutively ac- ing ultimately to cell growth, cell death, cell differentiation, cell tive form, G12V [12]. Subsequently, we showed that these pro- migration and invasion. files are also observed in binding of Rho by the other homologous BCH domains of BNIP-S [33], BNIP-XL [18] and 2. BCH domain as an emerging regulatory scaffold domain Cdc42GAP [19] and are distinct from that conferred by the GTPase-binding domains of GEFs and GAPs [14,15,20]. Further- 2.1. Discovery of the prototypical BCH domain from BNIP-2: from more, BNIP-2 appeared to promote the inactivation of Cdc42 receptor tyrosine kinase to small GTPase, MAP kinase and beyond by stimulating the apparent GTPase activity of Cdc42 in vitro and such effect is abrogated upon binding by FGFR [9] or nega- To understand the fundamental mechanism in cell signaling it is tively regulated by the homophilic interaction of BNIP-2 or its important to identify what and how one particular signaling pro- heterophilic interaction with Cdc42GAP [11]. Since then, by tein forms its protein interaction networks and how this process using yeast 2-hybrid, proteomics-based mass spectrometry and is regulated in vitro and in vivo. In our effort to decipher how acti- candidate approaches, we and others have expanded the family vation of the fibroblast growth factor receptor (FGFR) tyrosine ki- of BNIP-2 interactomes. We have further delineated how some nase could lead to novel signaling pathway, yeast 2-hybrid was of these BCH domains could in fact either activate or inactivate employed and BNIP-2 was identified as a candidate partner for specific Rho GTPases signaling depending on various cellular the cytoplasmic tail of the receptor [9]. Interestingly, only the ki- contexts the proteins are present, and they could directly or nase-dead mutants of the FGFR could ‘‘trap’’ and co-immunopre-
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