Universität zu Köln

Kohli, Priyanka ORCID: 0000-0002-0651-4008, Hoehne, Martin, Juengst, Christian, Bertsch, Sabine, Ebert, Lena K., Schauss, Astrid C., Benzing, Thomas, Rinschen, Markus M. and Schermer, Bernhard ORCID: 0000-0002-5194-9000 (2017). The ciliary membrane-associated proteome reveals actin-binding proteins as key components of cilia. EMBO Rep., 18 (9). S. 1521 - 1536. HOBOKEN: WILEY. ISSN 1469-3178

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Abstract

Primary cilia are sensory, antennae-like organelles present on the surface of many cell types. They have been involved in a variety of diseases collectively termed ciliopathies. As cilia are essential regulators of cell signaling, the composition of the ciliary membrane needs to be strictly regulated. To understand regulatory processes at the ciliary membrane, we report the targeting of a genetically engineered enzyme specifically to the ciliary membrane to allow biotinylation and identification of the membrane-associated proteome. Bioinformatic analysis of the comprehensive dataset reveals high-stoichiometric presence of actin-binding proteins inside the cilium. Immunofluorescence stainings and complementary interaction proteomic analyses confirm these findings. Depolymerization of branched F-actin causes further enrichment of the actin-binding and actin-related proteins in cilia, including Myosin 5a (Myo5a). Interestingly, Myo5a knockout decreases ciliation while enhanced levels of Myo5a are observed in cilia upon induction of ciliary disassembly. In summary, we present a novel approach to investigate dynamics of the ciliary membrane proteome in mammalian cells and identify actin-binding proteins as mechanosensitive components of cilia that might have important functions in cilia membrane dynamics.

Item Type:	Journal Article
Creators:	Creators Email ORCID ORCID Put Code Kohli, Priyanka UNSPECIFIED orcid.org/0000-0002-0651-4008 UNSPECIFIED Hoehne, Martin UNSPECIFIED UNSPECIFIED UNSPECIFIED Juengst, Christian UNSPECIFIED UNSPECIFIED UNSPECIFIED Bertsch, Sabine UNSPECIFIED UNSPECIFIED UNSPECIFIED Ebert, Lena K. UNSPECIFIED UNSPECIFIED UNSPECIFIED Schauss, Astrid C. UNSPECIFIED UNSPECIFIED UNSPECIFIED Benzing, Thomas UNSPECIFIED UNSPECIFIED UNSPECIFIED Rinschen, Markus M. UNSPECIFIED UNSPECIFIED UNSPECIFIED Schermer, Bernhard UNSPECIFIED orcid.org/0000-0002-5194-9000 UNSPECIFIED
URN:	urn:nbn:de:hbz:38-220215
DOI:	10.15252/embr.201643846
Journal or Publication Title:	EMBO Rep.
Volume:	18
Number:	9
Page Range:	S. 1521 - 1536
Date:	2017
Publisher:	WILEY
Place of Publication:	HOBOKEN
ISSN:	1469-3178
Language:	English
Faculty:	Unspecified
Divisions:	Unspecified
Subjects:	no entry
Uncontrolled Keywords:	Keywords Language VERTEBRATE PRIMARY CILIUM; NEGATIVE REGULATOR; CELL-CYCLE; CILIOGENESIS; TRANSPORT; LOCALIZATION; ENRICHMENT; IDENTIFICATION; PURIFICATION; MECHANISMS Multiple languages Biochemistry & Molecular Biology; Cell Biology Multiple languages
Refereed:	Yes
URI:	http://kups.ub.uni-koeln.de/id/eprint/22021