Primary cilia membrane assembly is initiated by Rab11 and transport protein particle II (TRAPPII) complex-dependent trafficking of Rabin8 to the centrosome

Christopher J Westlake; Lisa M Baye; Maxence V Nachury; Kevin J Wright; Karen E Ervin; Lilian Phu; Cecile Chalouni; John S Beck; Donald S Kirkpatrick; Diane C Slusarski; Val C Sheffield; Richard H Scheller; Peter K Jackson

doi:10.1073/pnas.1018823108

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Primary cilia membrane assembly is initiated by Rab11 and transport protein particle II (TRAPPII) complex-dependent trafficking of Rabin8 to the centrosome

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Primary cilia membrane assembly is initiated by Rab11 and transport protein particle II (TRAPPII) complex-dependent trafficking of Rabin8 to the centrosome

Christopher J Westlake, Lisa M Baye, Maxence V Nachury, Kevin J Wright, Karen E Ervin, Lilian Phu, Cecile Chalouni, John S Beck, Donald S Kirkpatrick, Diane C Slusarski, …

Proceedings of the National Academy of Sciences - PNAS, Vol.108(7), pp.2759-2764

02/15/2011

DOI: 10.1073/pnas.1018823108

PMCID: PMC3041065

PMID: 21273506

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Abstract

Sensory and signaling pathways are exquisitely organized in primary cilia. Bardet-Biedl syndrome (BBS) patients have compromised cilia and signaling. BBS proteins form the BBSome, which binds Rabin8, a guanine nucleotide exchange factor (GEF) activating the Rab8 GTPase, required for ciliary assembly. We now describe serum-regulated upstream vesicular transport events leading to centrosomal Rab8 activation and ciliary membrane formation. Using live microscopy imaging, we show that upon serum withdrawal Rab8 is observed to assemble the ciliary membrane in ∼100 min. Rab8-dependent ciliary assembly is initiated by the relocalization of Rabin8 to Rab11-positive vesicles that are transported to the centrosome. After ciliogenesis, Rab8 ciliary transport is strongly reduced, and this reduction appears to be associated with decreased Rabin8 centrosomal accumulation. Rab11-GTP associates with the Rabin8 COOH-terminal region and is required for Rabin8 preciliary membrane trafficking to the centrosome and for ciliogenesis. Using zebrafish as a model organism, we show that Rabin8 and Rab11 are associated with the BBS pathway. Finally, using tandem affinity purification and mass spectrometry, we determined that the transport protein particle (TRAPP) II complex associates with the Rabin8 NH(2)-terminal domain and show that TRAPP II subunits colocalize with centrosomal Rabin8 and are required for Rabin8 preciliary targeting and ciliogenesis.

Cilia - physiology

rab GTP-Binding Proteins - metabolism

Bardet-Biedl Syndrome - metabolism

Humans

Zebrafish

Bardet-Biedl Syndrome - physiopathology

Time-Lapse Imaging

Centrosome - metabolism

Two-Hybrid System Techniques

Animals

Carrier Proteins - metabolism

Transfection

Analysis of Variance

Guanine Nucleotide Exchange Factors - metabolism

Mass Spectrometry

Fluorescent Antibody Technique

Membranes - growth & development

Signal Transduction - physiology

Details

Title: Subtitle: Primary cilia membrane assembly is initiated by Rab11 and transport protein particle II (TRAPPII) complex-dependent trafficking of Rabin8 to the centrosome
Creators: Christopher J Westlake - Genentech, South San Francisco, CA 94080, USA
Lisa M Baye
Maxence V Nachury
Kevin J Wright
Karen E Ervin
Lilian Phu
Cecile Chalouni
John S Beck
Donald S Kirkpatrick
Diane C Slusarski
Val C Sheffield
Richard H Scheller
Peter K Jackson
Resource Type: Journal article
Publication Details: Proceedings of the National Academy of Sciences - PNAS, Vol.108(7), pp.2759-2764
DOI: 10.1073/pnas.1018823108
PMID: 21273506
PMCID: PMC3041065
NLM abbreviation: Proc Natl Acad Sci U S A
ISSN: 0027-8424
eISSN: 1091-6490
Publisher: United States
Grant note: R01 EY011298 / NEI NIH HHS T32 HL007121 / NHLBI NIH HHS Howard Hughes Medical Institute CA112369 / NCI NIH HHS R01 GM089933 / NIGMS NIH HHS R01 EY017168 / NEI NIH HHS EY017168 / NEI NIH HHS R01EY011298 / NEI NIH HHS R01 CA112369 / NCI NIH HHS
Language: English
Date published: 02/15/2011
Academic Unit: Stead Family Department of Pediatrics; Iowa Neuroscience Institute; Medical Genetics and Genomics; Biology; Ophthalmology and Visual Sciences
Record Identifier: 9983992063002771

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