Biochemical and structural insights into Rab12 interactions with RILP and its family members

Abstract Alongside its biosynthetic functions, the small GTPase Rab12 negatively regulates mast cell (MC) exocytosis by its interaction with RILP to promote retrograde transport of the MC secretory granules. Given the role of Rab effectors in mediating Rab functions, in this study we used biochemica...

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Autores principales:	Jana Omar, Efrat Rosenbaum, Adi Efergan, Bayan Abu Sneineh, Adva Yeheskel, Yuto Maruta, Mitsunori Fukuda, Ronit Sagi-Eisenberg
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2021
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/421863333db34649b334e57221653e1f
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spelling	oai:doaj.org-article:421863333db34649b334e57221653e1f2021-12-02T15:54:59ZBiochemical and structural insights into Rab12 interactions with RILP and its family members10.1038/s41598-021-89394-y2045-2322https://doaj.org/article/421863333db34649b334e57221653e1f2021-05-01T00:00:00Zhttps://doi.org/10.1038/s41598-021-89394-yhttps://doaj.org/toc/2045-2322Abstract Alongside its biosynthetic functions, the small GTPase Rab12 negatively regulates mast cell (MC) exocytosis by its interaction with RILP to promote retrograde transport of the MC secretory granules. Given the role of Rab effectors in mediating Rab functions, in this study we used biochemical and in silico tools to decipher Rab12 interactions with its RILP family effectors. We show that Rab12 interacts with RILP, RILP-L1 and RILP-L2 independently of each other, whereby lysine-71, in mouse Rab12, is critical for Rab12 interactions with RILP-L1 or RILP-L2, but is dispensable for the binding of RILP. Focusing on RILP, and relying on molecular dynamics simulations, functional mutational analyses and peptide inhibition assays, we propose a model for the Rab12-RILP complex, consisting of a RILP homodimer and a single molecule of active Rab12, that interacts with the RILP homology domain (RHD) of one RILP monomer and a C-terminal threonine in the other monomer via its switch I and switch II regions. Mutational analyses of RILP RHD also demonstrate its involvement in the regulation of MC secretory granule transport. Jointly, our results provide structural and functional insights into the Rab12-RILP complex on the basis of which new tools could be generated for decoding Rab12 functions.Jana OmarEfrat RosenbaumAdi EferganBayan Abu SneinehAdva YeheskelYuto MarutaMitsunori FukudaRonit Sagi-EisenbergNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 11, Iss 1, Pp 1-16 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Jana Omar Efrat Rosenbaum Adi Efergan Bayan Abu Sneineh Adva Yeheskel Yuto Maruta Mitsunori Fukuda Ronit Sagi-Eisenberg Biochemical and structural insights into Rab12 interactions with RILP and its family members
description	Abstract Alongside its biosynthetic functions, the small GTPase Rab12 negatively regulates mast cell (MC) exocytosis by its interaction with RILP to promote retrograde transport of the MC secretory granules. Given the role of Rab effectors in mediating Rab functions, in this study we used biochemical and in silico tools to decipher Rab12 interactions with its RILP family effectors. We show that Rab12 interacts with RILP, RILP-L1 and RILP-L2 independently of each other, whereby lysine-71, in mouse Rab12, is critical for Rab12 interactions with RILP-L1 or RILP-L2, but is dispensable for the binding of RILP. Focusing on RILP, and relying on molecular dynamics simulations, functional mutational analyses and peptide inhibition assays, we propose a model for the Rab12-RILP complex, consisting of a RILP homodimer and a single molecule of active Rab12, that interacts with the RILP homology domain (RHD) of one RILP monomer and a C-terminal threonine in the other monomer via its switch I and switch II regions. Mutational analyses of RILP RHD also demonstrate its involvement in the regulation of MC secretory granule transport. Jointly, our results provide structural and functional insights into the Rab12-RILP complex on the basis of which new tools could be generated for decoding Rab12 functions.
format	article
author	Jana Omar Efrat Rosenbaum Adi Efergan Bayan Abu Sneineh Adva Yeheskel Yuto Maruta Mitsunori Fukuda Ronit Sagi-Eisenberg
author_facet	Jana Omar Efrat Rosenbaum Adi Efergan Bayan Abu Sneineh Adva Yeheskel Yuto Maruta Mitsunori Fukuda Ronit Sagi-Eisenberg
author_sort	Jana Omar
title	Biochemical and structural insights into Rab12 interactions with RILP and its family members
title_short	Biochemical and structural insights into Rab12 interactions with RILP and its family members
title_full	Biochemical and structural insights into Rab12 interactions with RILP and its family members
title_fullStr	Biochemical and structural insights into Rab12 interactions with RILP and its family members
title_full_unstemmed	Biochemical and structural insights into Rab12 interactions with RILP and its family members
title_sort	biochemical and structural insights into rab12 interactions with rilp and its family members
publisher	Nature Portfolio
publishDate	2021
url	https://doaj.org/article/421863333db34649b334e57221653e1f
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Biochemical and structural insights into Rab12 interactions with RILP and its family members

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