Three-dimensional structure of the Ras-interacting domain of RalGDS

Huang, Li-Shar; X, Weng; Hofer, Franz; Gs, Martin; Sh, Kim

doi:10.1038/nsb0897-609

Cited by 79 publications

(69 citation statements)

References 41 publications

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“…The ability of RA domains to bind Ras-like small GTPases appears to have been acquired subsequent to the emergence of this domain, and it is possible that those RA domains that have acquired the ability to bind Ras-like GTPases may yet also retain the ability to bind another class of partners, presumably related to those that bind the RASSF1 RA domain. It is important to note that the primary sequence of the Ras-binding domain of the Raf kinase does not conform to the RA motif, suggesting that the ability to bind Ras-like Smgs evolved independently, but in a convergent manner; in spite of their lack of sequence similarity, the tertiary structure of the Raf RBD (Nassar et al, 1995) and the Ral GDS RA domain (Huang et al, 1997) exhibit a similar ubiquitin superfold. The diversity in Ras-binding motifs is greater yet, as the aminoterminus of the Byr2 kinase binds Ras-GTP speci®cally (Akasaka et al, 1996) but shares no resemblance in primary sequence to either the Raf RBD or RA motifs.…”

Section: Discussionmentioning

confidence: 99%

The putative tumor suppressor RASSF1A homodimerizes and heterodimerizes with the Ras-GTP binding protein Nore1

et al. 2002

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Nore and RASSF1A are noncatalytic proteins that share 50% identity over their carboxyterminal 300 AA, a segment that encompasses a putative Ras-Rap association (RA) domain. RASSF1 is expressed as several splice variants, each of which contain an RA domain, however the 340 AA RASSF1A, but not the shorter RASSF1C variant, is a putative tumor suppressor. Nore binds to Ras and several Ras-like GTPases in a GTP dependent fashion however neither RASSF1 (A or C) or the C. elegans Nore/RASSF1 homolog, T24F1.3 exhibit any interaction with Ras or six other Ras-like GTPases in a yeast two-hybrid expression assay. A low recovery of RASSF1A (but not RASSF1C) in association with RasG12V is observed however on transient expression in COS cells. Nore and RASSF1A can each e ciently homodimerize and heterodimerize with each other through their nonhomologous aminoterminal segments. Recombinant RASSF1C exhibits a much weaker ability to homodimerize or heterodimerize; thus the binding of RASSF1C to Nore is very much less than the binding of RASSF1A to Nore. The association of RASSF1A with RasG12V in COS cells appears to re¯ect the heterodimerization of RASSF1A with Nore, inasmuch the recovery of RASSF1A with RasG12V is increased by concurrent expression of full length Nore, and abolished by expression of Nore deleted of its RA domain. The preferential ability of RASSF1A to heterodimerize with Nore and thereby associate with Ras-like GTPases may be relevant to its putative tumor suppressor function.

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Section: Discussionmentioning

confidence: 99%

The putative tumor suppressor RASSF1A homodimerizes and heterodimerizes with the Ras-GTP binding protein Nore1

et al. 2002

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“…In fact, NMR chemical shift mapping studies of RalGDS-RID in a complex with Ras, indicate an interaction similar to the intermolecular beta-sheet observed for the complex between Ras and Raf-RBS (Geyer et al, 1997). Furthermore, mutagenesis studies indicate that several residues in RalGDS-RID that mediate binding to Ras are located at sites that correpond to those important for binding by Raf-RBS (Huang et al, 1997;Geyer et al, 1997). Hence, it is tempting to speculate, given the structural and mutagenesis data, that the RalGDS-Ras complex will be similar to the Ras-Raf complex, and may provide a common structural basis for interaction with a subset of Ras e ectors.…”

Section: Ras Mediates Its Actions Through Interaction With Multiple Ementioning

confidence: 78%

“…The overall topology of Ras GAP-CAT, in turn, is quite distinct from that of Raf-RBS (Emerson et al, 1995;Nassar et al, 1995), in that Raf-RBS possesses a mixed a/b fold similar to that of ubiquitin. Interestingly, despite the low level of sequence homology between Raf-RBS and the Ras interaction domain of RalGDS (RalGDS-RID), Raf-RBS has been shown to have a similar tertiary fold to RalGDS-RID (Huang et al, 1997;Geyer et al, 1997). In fact, NMR chemical shift mapping studies of RalGDS-RID in a complex with Ras, indicate an interaction similar to the intermolecular beta-sheet observed for the complex between Ras and Raf-RBS (Geyer et al, 1997).…”

Section: Ras Mediates Its Actions Through Interaction With Multiple Ementioning

confidence: 99%

Increasing complexity of Ras signaling

et al. 1998

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The initial discovery that ras genes endowed retroviruses with potent carcinogenic properties and the subsequent determination that mutated ras genes were present in a wide variety of human cancers, prompted a strong suspicion that the growth-promoting actions of mutated Ras proteins contribute to their aberrant regulation of growth stimulatory signaling pathways. In 1993, a remarkable convergence of experimental observations from genetic analyses of Drosophila, S. cerevisiae and C. elegans as well as biochemical and biological studies in mammalian cells came together to de®ne a clear role for Ras in signal transduction. What emerged was an elegant linear signaling pathway where Ras functions as a relay switch that is positioned downstream of cell surface receptor tyrosine kinases and upstream of a cytoplasmic cascade of kinases that included the mitogen-activated protein kinases (MAPKs). Activated MAPKs in turn regulated the activities of nuclear transcription factors. Thus, a signaling cascade where every component between the cell surface and the nucleus was de®ned and conserved in worms,¯ies and man. This was a remarkable achievement in our e orts to appreciate how the aberrant function of Ras proteins may contribute to the malignant growth properties of the cancer cell. However, the identi®cation of this pathway has proven to be just the beginning, rather than the culmination, of our understanding of Ras in signal transduction. Instead, we now appreciate that this simple linear pathway represents but a minor component of a very complex signaling circuitry. Ras signaling has emerged to involve a complex array of signaling pathways, where cross-talk, feedback loops, branch points and multi-component signaling complexes are recurring themes. The simplest concept of a signaling cascade, where each component simply relays the same message to the next, is clearly not the case. In this review, we summarize our current understanding of Ras signal transduction with an emphasis on new complexities associated with the recognition and/or activation of cellular e ectors, and the diverse array of signaling pathways mediated by interaction between Ras and Ras-subfamily proteins with multiple e ectors.

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“…The fraction of this region provided by ␤-strand 2 as well as part of ␤-strand 1 and potentially the C-terminal end of ␣-helix 1 may, as has been observed for the RalGDS (Fig. 3B) and the PI3K Ras-binding domain (31)(32)(33), mediate the interaction with complementary negatively charged patches in the switch I and II regions of a regulatory GTPase. The F0 domain of the protein talin, an actinbinding protein involved in integrin activation, also shows a ubiquitin-like fold, including a similar although smaller positive surface patch.…”

Section: Resultsmentioning

confidence: 99%

Structure, Dynamics, Lipid Binding, and Physiological Relevance of the Putative GTPase-binding Domain of Dictyostelium Formin C

Dames

Junemann

Sass

et al. 2011

Journal of Biological Chemistry

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Dictyostelium Formin C (ForC) is involved in the regulation of local actin cytoskeleton reorganization (e.g. during cellular adhesion or migration). ForC contains formin homology 2 and 3 (FH2 and -3) domains and an N-terminal putative GTPase-binding domain (GBD) but lacks a canonical FH1 region. To better understand the role of the GBD, its structure, dynamics, lipid-binding properties, and cellular functions were analyzed by NMR and CD spectroscopy and by in vivo fluorescence microscopy. Moreover, the program CS-Rosetta was tested for the structure prediction based on chemical shift data only. The ForC GBD adopts an ubiquitinlike ␣/␤-roll fold with an unusually long loop between ␤-strands 1 and 2. Based on the lipid-binding data, the presence of DPC micelles induces the formation of ␣-helical secondary structure and a rearrangement of the tertiary structure. Lipid-binding studies with a mutant protein and a peptide suggest that the ␤1-␤2 loop is not relevant for these conformational changes. Whereas small amounts of negatively charged phosphoinositides (1,2-dioctanoyl-sn-glycero-3-(phosphoinositol 4,5-bisphosphate) and 1,2-dihexanoyl-sn-glycero-3-(phosphoinositol 3,4,5-trisphosphate)) lower the micelle concentration necessary to induce the observed spectral changes, other negatively charged phospholipids (1,2-dihexanoyl-sn-glycero-3-(phospho-L-serine) and 1,2-dihexanoyl-snglycero-3-phospho-(1 -rac-glycerol)) had no such effect. Interestingly, bicelles and micelles composed of diacylphosphocholines had no effect on the GBD structure. Our data suggest a model in which part of the large positively charged surface area of the GBD mediates localization to specific membrane patches, thereby regulating interactions with signaling proteins. Our cellular localization studies show that both the GBD and the FH3 domain are required for ForC targeting to cell-cell contacts and early phagocytic cups and macropinosomes.Formins are widely expressed multidomain proteins that regulate the spatial and temporal organization of the actin and microtubule cytoskeleton, thereby affecting important cellular functions, such as cytokinesis and cell-cell adhesion (1-3). The defining element is the ϳ400-amino acid-encompassing "formin homology 2" (FH2) 2 domain that can tightly associate with the barbed end of elongating actin filaments (4). In nearly all formins, the FH2 domain is preceded by a prolinerich "formin homology 1" (FH1) region that varies in length and the number of binding sites for profilin and that is used to recruit profilin-actin complexes for filament elongation. The influence of the FH2 domain on the rate of filament elongation depends on the length and composition of the FH1 region and the connecting linker (2, 4, 5). Most formins contain additional regulatory domains. The N-terminal "formin homology 3" (FH3) domain mediates interactions with autoinhibitory regulatory elements in the C terminus as well as with other formin regulators. The FH3 domain can be further divided into functional subdomains, such as an armadill...

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Three-dimensional structure of the Ras-interacting domain of RalGDS

Cited by 79 publications

References 41 publications

The putative tumor suppressor RASSF1A homodimerizes and heterodimerizes with the Ras-GTP binding protein Nore1

The putative tumor suppressor RASSF1A homodimerizes and heterodimerizes with the Ras-GTP binding protein Nore1

Increasing complexity of Ras signaling

Structure, Dynamics, Lipid Binding, and Physiological Relevance of the Putative GTPase-binding Domain of Dictyostelium Formin C

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