Identification of the centrosomal maturation factor SSX2IP as a Wtip-binding partner by targeted proximity biotinylation.

Wilms tumor-1-interacting protein (Wtip) is a LIM-domain-containing adaptor that links cell junctions with actomyosin complexes and modulates actomyosin contractility and ciliogenesis in Xenopus embryos. The Wtip C-terminus with three LIM domains associates with the actin-binding protein Shroom3 and...

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Autores principales:	Alice H Reis, Bo Xiang, Olga Ossipova, Keiji Itoh, Sergei Y Sokol
Formato:	article
Lenguaje:	EN
Publicado:	Public Library of Science (PLoS) 2021
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/1fdcc85544364d618821a886b02d0c14
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spelling	oai:doaj.org-article:1fdcc85544364d618821a886b02d0c142021-12-02T20:16:29ZIdentification of the centrosomal maturation factor SSX2IP as a Wtip-binding partner by targeted proximity biotinylation.1932-620310.1371/journal.pone.0259068https://doaj.org/article/1fdcc85544364d618821a886b02d0c142021-01-01T00:00:00Zhttps://doi.org/10.1371/journal.pone.0259068https://doaj.org/toc/1932-6203Wilms tumor-1-interacting protein (Wtip) is a LIM-domain-containing adaptor that links cell junctions with actomyosin complexes and modulates actomyosin contractility and ciliogenesis in Xenopus embryos. The Wtip C-terminus with three LIM domains associates with the actin-binding protein Shroom3 and modulates Shroom3-induced apical constriction in ectoderm cells. By contrast, the N-terminal domain localizes to apical junctions in the ectoderm and basal bodies in skin multiciliated cells, but its interacting partners remain largely unknown. Targeted proximity biotinylation (TPB) using anti-GFP antibody fused to the biotin ligase BirA identified SSX2IP as a candidate protein that binds GFP-WtipN. SSX2IP, also known as Msd1 or ADIP, is a component of cell junctions, centriolar satellite protein and a targeting factor for ciliary membrane proteins. WtipN physically associated with SSX2IP and the two proteins readily formed mixed aggregates in overexpressing cells. By contrast, we observed only partial colocalization of full length Wtip and SSX2IP, suggesting that Wtip adopts a 'closed' conformation in the cell. Furthermore, the double depletion of Wtip and SSX2IP in early embryos uncovered the functional interaction of the two proteins during neural tube closure. Our results suggest that the association of SSX2IP and Wtip is essential for cell junction remodeling and morphogenetic processes that accompany neurulation. We propose that TPB can be a general approach that is applicable to other GFP-tagged proteins.Alice H ReisBo XiangOlga OssipovaKeiji ItohSergei Y SokolPublic Library of Science (PLoS)articleMedicineRScienceQENPLoS ONE, Vol 16, Iss 10, p e0259068 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Alice H Reis Bo Xiang Olga Ossipova Keiji Itoh Sergei Y Sokol Identification of the centrosomal maturation factor SSX2IP as a Wtip-binding partner by targeted proximity biotinylation.
description	Wilms tumor-1-interacting protein (Wtip) is a LIM-domain-containing adaptor that links cell junctions with actomyosin complexes and modulates actomyosin contractility and ciliogenesis in Xenopus embryos. The Wtip C-terminus with three LIM domains associates with the actin-binding protein Shroom3 and modulates Shroom3-induced apical constriction in ectoderm cells. By contrast, the N-terminal domain localizes to apical junctions in the ectoderm and basal bodies in skin multiciliated cells, but its interacting partners remain largely unknown. Targeted proximity biotinylation (TPB) using anti-GFP antibody fused to the biotin ligase BirA identified SSX2IP as a candidate protein that binds GFP-WtipN. SSX2IP, also known as Msd1 or ADIP, is a component of cell junctions, centriolar satellite protein and a targeting factor for ciliary membrane proteins. WtipN physically associated with SSX2IP and the two proteins readily formed mixed aggregates in overexpressing cells. By contrast, we observed only partial colocalization of full length Wtip and SSX2IP, suggesting that Wtip adopts a 'closed' conformation in the cell. Furthermore, the double depletion of Wtip and SSX2IP in early embryos uncovered the functional interaction of the two proteins during neural tube closure. Our results suggest that the association of SSX2IP and Wtip is essential for cell junction remodeling and morphogenetic processes that accompany neurulation. We propose that TPB can be a general approach that is applicable to other GFP-tagged proteins.
format	article
author	Alice H Reis Bo Xiang Olga Ossipova Keiji Itoh Sergei Y Sokol
author_facet	Alice H Reis Bo Xiang Olga Ossipova Keiji Itoh Sergei Y Sokol
author_sort	Alice H Reis
title	Identification of the centrosomal maturation factor SSX2IP as a Wtip-binding partner by targeted proximity biotinylation.
title_short	Identification of the centrosomal maturation factor SSX2IP as a Wtip-binding partner by targeted proximity biotinylation.
title_full	Identification of the centrosomal maturation factor SSX2IP as a Wtip-binding partner by targeted proximity biotinylation.
title_fullStr	Identification of the centrosomal maturation factor SSX2IP as a Wtip-binding partner by targeted proximity biotinylation.
title_full_unstemmed	Identification of the centrosomal maturation factor SSX2IP as a Wtip-binding partner by targeted proximity biotinylation.
title_sort	identification of the centrosomal maturation factor ssx2ip as a wtip-binding partner by targeted proximity biotinylation.
publisher	Public Library of Science (PLoS)
publishDate	2021
url	https://doaj.org/article/1fdcc85544364d618821a886b02d0c14
work_keys_str_mv	AT alicehreis identificationofthecentrosomalmaturationfactorssx2ipasawtipbindingpartnerbytargetedproximitybiotinylation AT boxiang identificationofthecentrosomalmaturationfactorssx2ipasawtipbindingpartnerbytargetedproximitybiotinylation AT olgaossipova identificationofthecentrosomalmaturationfactorssx2ipasawtipbindingpartnerbytargetedproximitybiotinylation AT keijiitoh identificationofthecentrosomalmaturationfactorssx2ipasawtipbindingpartnerbytargetedproximitybiotinylation AT sergeiysokol identificationofthecentrosomalmaturationfactorssx2ipasawtipbindingpartnerbytargetedproximitybiotinylation
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Identification of the centrosomal maturation factor SSX2IP as a Wtip-binding partner by targeted proximity biotinylation.

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