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

*Corresponding author for this work

Research output: Contribution to journalJournal articleResearchpeer-review

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
Volume20
Issue number10
Number of pages23
ISSN1469-221X
DOIs
Publication statusPublished - 4. Oct 2019

Fingerprint

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

Keywords

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

Cite this

Kuhns, S., Seixas, C., Pestana, S., Tavares, B., Nogueira, R., Jacinto, R., ... Blacque, O. E. (2019). Rab35 controls cilium length, function and membrane composition. EMBO Reports, 20(10), [e47625]. https://doi.org/10.15252/embr.201847625
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 ; Vol. 20, No. 10.
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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.",
keywords = "Arl13b, cilia, left-right asymmetry, Rab35, Smoothened",
author = "Stefanie Kuhns and Cec{\'i}lia Seixas and Sara Pestana and B{\'a}rbara Tavares and Renata Nogueira and Raquel Jacinto and Ramalho, {Jos{\'e} S.} and Simpson, {Jeremy C.} and Andersen, {Jens S.} and Arnaud Echard and Lopes, {Susana S.} and Barral, {Duarte C.} and Blacque, {Oliver E.}",
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month = "10",
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Kuhns, S, Seixas, C, Pestana, S, Tavares, B, Nogueira, R, Jacinto, R, Ramalho, JS, Simpson, JC, Andersen, JS, Echard, A, Lopes, SS, Barral, DC & Blacque, OE 2019, 'Rab35 controls cilium length, function and membrane composition', EMBO Reports, vol. 20, no. 10, e47625. https://doi.org/10.15252/embr.201847625

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, Vol. 20, No. 10, e47625, 04.10.2019.

Research output: Contribution to journalJournal articleResearchpeer-review

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 - 2019/10/4

Y1 - 2019/10/4

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

KW - cilia

KW - left-right asymmetry

KW - Rab35

KW - Smoothened

U2 - 10.15252/embr.201847625

DO - 10.15252/embr.201847625

M3 - Journal article

C2 - 31432619

AN - SCOPUS:85070872369

VL - 20

JO - E M B O Reports

JF - E M B O Reports

SN - 1469-221X

IS - 10

M1 - e47625

ER -

Kuhns S, Seixas C, Pestana S, Tavares B, Nogueira R, Jacinto R et al. Rab35 controls cilium length, function and membrane composition. EMBO Reports. 2019 Oct 4;20(10). e47625. https://doi.org/10.15252/embr.201847625