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Characterisation of HRas local signal transduction networks using engineered site-specific exchange factors

AuthorsHerrero, Ana; Reis-Cardoso, Mariana; Jiménez Gómez, Iñaki; Doherty, Carolanne; Agudo-Ibáñez, Lorena; Pinto, Adán; Calvo, Fernando CSIC ORCID ; Kolch, Walter; Crespo, Piero CSIC ORCID; Matallanas, David
CDC25 domain
Signalling network
Issue Date2020
PublisherTaylor & Francis
CitationSmall GTPases 11(5): 371-383 (2020)
AbstractRas GTPases convey signals from different types of membranes. At these locations, different Ras isoforms, interactors and regulators generate different biochemical signals and biological outputs. The study of Ras localisation-specific signal transduction networks has been hampered by our inability to specifically activate each of these Ras pools. Here, we describe a new set of site-specific tethered exchange factors, engineered by fusing the RasGRF1 CDC25 domain to sub-localisation-defining cues, whereby Ras pools at specific locations can be precisely activated. We show that the CDC25 domain has a high specificity for activating HRas but not NRas and KRas. This unexpected finding means that our constructs mainly activate endogenous HRas. Hence, their use enabled us to identify distinct pathways regulated by HRas in endomembranes and plasma membrane microdomains. Importantly, these new constructs unveil different patterns of HRas activity specified by their subcellular localisation. Overall, the targeted GEFs described herein constitute ideal tools for dissecting spatially-defined HRas biochemical and biological functions.
Publisher version (URL)http://dx.doi.org/10.1080/21541248.2017.1406434
Appears in Collections:(IBBTEC) Artículos
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