Rab35 controls cilium length, function and membrane composition

Stefanie Kuhns, Cecília Seixas, Sara Pestana, Bárbara Tavares, Renata Nogueira, Raquel Jacinto, José S. Ramalho, Jeremy C. Simpson, Jens S. Andersen, Arnaud Echard, Susana S. Lopes, Duarte C. Barral, Oliver E. Blacque

Research output: Contribution to journalArticle

Abstract

Rab and Arl guanine nucleotide-binding (G) proteins regulate trafficking pathways essential for the formation, function and composition of primary cilia, which are sensory devices associated with Sonic hedgehog (Shh) signalling and ciliopathies. Here, using mammalian cells and zebrafish, we uncover ciliary functions for Rab35, a multitasking G protein with endocytic recycling, actin remodelling and cytokinesis roles. Rab35 loss via siRNAs, morpholinos or knockout reduces cilium length in mammalian cells and the zebrafish left-right organiser (Kupffer's vesicle) and causes motile cilia-associated left-right asymmetry defects. Consistent with these observations, GFP-Rab35 localises to cilia, as do GEF (DENND1B) and GAP (TBC1D10A) Rab35 regulators, which also regulate ciliary length and Rab35 ciliary localisation. Mammalian Rab35 also controls the ciliary membrane levels of Shh signalling regulators, promoting ciliary targeting of Smoothened, limiting ciliary accumulation of Arl13b and the inositol polyphosphate 5-phosphatase (INPP5E). Rab35 additionally regulates ciliary PI(4,5)P2 levels and interacts with Arl13b. Together, our findings demonstrate roles for Rab35 in regulating cilium length, function and membrane composition and implicate Rab35 in pathways controlling the ciliary levels of Shh signal regulators.

Original languageEnglish
Article numbere47625
JournalEmbo Reports
DOIs
Publication statusPublished - 1 Jan 2019

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Cilia
Hedgehogs
Membranes
GTP-Binding Proteins
Chemical analysis
Cells
Zebrafish
Morpholinos
Polyphosphates
Multitasking
Guanine Nucleotides
Inositol
Phosphoric Monoester Hydrolases
Recycling
Actins
Cytokinesis
Protein Transport
Defects
Carrier Proteins
Equipment and Supplies

Keywords

  • Arl13b
  • cilia
  • left-right asymmetry
  • Rab35
  • Smoothened

Cite this

Kuhns, Stefanie ; Seixas, Cecília ; Pestana, Sara ; Tavares, Bárbara ; Nogueira, Renata ; Jacinto, Raquel ; Ramalho, José S. ; Simpson, Jeremy C. ; Andersen, Jens S. ; Echard, Arnaud ; Lopes, Susana S. ; Barral, Duarte C. ; Blacque, Oliver E. / Rab35 controls cilium length, function and membrane composition. In: Embo Reports. 2019.
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Rab35 controls cilium length, function and membrane composition. / Kuhns, Stefanie; Seixas, Cecília; Pestana, Sara; Tavares, Bárbara; Nogueira, Renata; Jacinto, Raquel; Ramalho, José S.; Simpson, Jeremy C.; Andersen, Jens S.; Echard, Arnaud; Lopes, Susana S.; Barral, Duarte C.; Blacque, Oliver E.

In: Embo Reports, 01.01.2019.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Rab35 controls cilium length, function and membrane composition

AU - Kuhns, Stefanie

AU - Seixas, Cecília

AU - Pestana, Sara

AU - Tavares, Bárbara

AU - Nogueira, Renata

AU - Jacinto, Raquel

AU - Ramalho, José S.

AU - Simpson, Jeremy C.

AU - Andersen, Jens S.

AU - Echard, Arnaud

AU - Lopes, Susana S.

AU - Barral, Duarte C.

AU - Blacque, Oliver E.

PY - 2019/1/1

Y1 - 2019/1/1

N2 - Rab and Arl guanine nucleotide-binding (G) proteins regulate trafficking pathways essential for the formation, function and composition of primary cilia, which are sensory devices associated with Sonic hedgehog (Shh) signalling and ciliopathies. Here, using mammalian cells and zebrafish, we uncover ciliary functions for Rab35, a multitasking G protein with endocytic recycling, actin remodelling and cytokinesis roles. Rab35 loss via siRNAs, morpholinos or knockout reduces cilium length in mammalian cells and the zebrafish left-right organiser (Kupffer's vesicle) and causes motile cilia-associated left-right asymmetry defects. Consistent with these observations, GFP-Rab35 localises to cilia, as do GEF (DENND1B) and GAP (TBC1D10A) Rab35 regulators, which also regulate ciliary length and Rab35 ciliary localisation. Mammalian Rab35 also controls the ciliary membrane levels of Shh signalling regulators, promoting ciliary targeting of Smoothened, limiting ciliary accumulation of Arl13b and the inositol polyphosphate 5-phosphatase (INPP5E). Rab35 additionally regulates ciliary PI(4,5)P2 levels and interacts with Arl13b. Together, our findings demonstrate roles for Rab35 in regulating cilium length, function and membrane composition and implicate Rab35 in pathways controlling the ciliary levels of Shh signal regulators.

AB - Rab and Arl guanine nucleotide-binding (G) proteins regulate trafficking pathways essential for the formation, function and composition of primary cilia, which are sensory devices associated with Sonic hedgehog (Shh) signalling and ciliopathies. Here, using mammalian cells and zebrafish, we uncover ciliary functions for Rab35, a multitasking G protein with endocytic recycling, actin remodelling and cytokinesis roles. Rab35 loss via siRNAs, morpholinos or knockout reduces cilium length in mammalian cells and the zebrafish left-right organiser (Kupffer's vesicle) and causes motile cilia-associated left-right asymmetry defects. Consistent with these observations, GFP-Rab35 localises to cilia, as do GEF (DENND1B) and GAP (TBC1D10A) Rab35 regulators, which also regulate ciliary length and Rab35 ciliary localisation. Mammalian Rab35 also controls the ciliary membrane levels of Shh signalling regulators, promoting ciliary targeting of Smoothened, limiting ciliary accumulation of Arl13b and the inositol polyphosphate 5-phosphatase (INPP5E). Rab35 additionally regulates ciliary PI(4,5)P2 levels and interacts with Arl13b. Together, our findings demonstrate roles for Rab35 in regulating cilium length, function and membrane composition and implicate Rab35 in pathways controlling the ciliary levels of Shh signal regulators.

KW - Arl13b

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KW - left-right asymmetry

KW - Rab35

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DO - 10.15252/embr.201847625

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