Binding of IFT22 to the intraflagellar transport complex is essential for flagellum assembly

Stefanie Wachter; Jamin Jung; Shahaan Shafiq; Jerome Basquin; Cécile Fort; Philippe Bastin; Esben Lorentzen

doi:10.15252/embj.2018101251

Binding of IFT22 to the intraflagellar transport complex is essential for flagellum assembly

Stefanie Wachter, Jamin Jung, Shahaan Shafiq, Jerome Basquin, Cécile Fort, Philippe Bastin, Esben Lorentzen^*

^*Corresponding author for this work

Research output: Contribution to journal/Conference contribution in journal/Contribution to newspaper › Journal article › Research › peer-review

Abstract

Intraflagellar transport (IFT) relies on motor proteins and the IFT complex to construct cilia and flagella. The IFT complex subunit IFT22/RabL5 has sequence similarity with small GTPases although the nucleotide specificity is unclear because of non-conserved G4/G5 motifs. We show that IFT22 specifically associates with G-nucleotides and present crystal structures of IFT22 in complex with GDP, GTP, and with IFT74/81. Our structural analysis unravels an unusual GTP/GDP-binding mode of IFT22 bypassing the classical G4 motif. The GTPase switch regions of IFT22 become ordered upon complex formation with IFT74/81 and mediate most of the IFT22-74/81 interactions. Structure-based mutagenesis reveals that association of IFT22 with the IFT complex is essential for flagellum construction in Trypanosoma brucei although IFT22 GTP-loading is not strictly required.

Original language	English
Article number	e101251
Journal	EMBO Journal
Volume	38
Issue	9
ISSN	0261-4189
DOIs	https://doi.org/10.15252/embj.2018101251
Publication status	Published - May 2019

Keywords

cilia
GTPase
IFT22
intraflagellar transport
Trypanosoma brucei

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https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6484408/pdf/EMBJ-38-e101251.pdf

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title = "Binding of IFT22 to the intraflagellar transport complex is essential for flagellum assembly",

abstract = "Intraflagellar transport (IFT) relies on motor proteins and the IFT complex to construct cilia and flagella. The IFT complex subunit IFT22/RabL5 has sequence similarity with small GTPases although the nucleotide specificity is unclear because of non-conserved G4/G5 motifs. We show that IFT22 specifically associates with G-nucleotides and present crystal structures of IFT22 in complex with GDP, GTP, and with IFT74/81. Our structural analysis unravels an unusual GTP/GDP-binding mode of IFT22 bypassing the classical G4 motif. The GTPase switch regions of IFT22 become ordered upon complex formation with IFT74/81 and mediate most of the IFT22-74/81 interactions. Structure-based mutagenesis reveals that association of IFT22 with the IFT complex is essential for flagellum construction in Trypanosoma brucei although IFT22 GTP-loading is not strictly required.",

keywords = "cilia, GTPase, IFT22, intraflagellar transport, Trypanosoma brucei",

author = "Stefanie Wachter and Jamin Jung and Shahaan Shafiq and Jerome Basquin and C{\'e}cile Fort and Philippe Bastin and Esben Lorentzen",

year = "2019",

month = may,

doi = "10.15252/embj.2018101251",

language = "English",

volume = "38",

journal = "EMBO Journal",

issn = "0261-4189",

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T1 - Binding of IFT22 to the intraflagellar transport complex is essential for flagellum assembly

AU - Wachter, Stefanie

AU - Jung, Jamin

AU - Shafiq, Shahaan

AU - Basquin, Jerome

AU - Fort, Cécile

AU - Bastin, Philippe

AU - Lorentzen, Esben

PY - 2019/5

Y1 - 2019/5

N2 - Intraflagellar transport (IFT) relies on motor proteins and the IFT complex to construct cilia and flagella. The IFT complex subunit IFT22/RabL5 has sequence similarity with small GTPases although the nucleotide specificity is unclear because of non-conserved G4/G5 motifs. We show that IFT22 specifically associates with G-nucleotides and present crystal structures of IFT22 in complex with GDP, GTP, and with IFT74/81. Our structural analysis unravels an unusual GTP/GDP-binding mode of IFT22 bypassing the classical G4 motif. The GTPase switch regions of IFT22 become ordered upon complex formation with IFT74/81 and mediate most of the IFT22-74/81 interactions. Structure-based mutagenesis reveals that association of IFT22 with the IFT complex is essential for flagellum construction in Trypanosoma brucei although IFT22 GTP-loading is not strictly required.

AB - Intraflagellar transport (IFT) relies on motor proteins and the IFT complex to construct cilia and flagella. The IFT complex subunit IFT22/RabL5 has sequence similarity with small GTPases although the nucleotide specificity is unclear because of non-conserved G4/G5 motifs. We show that IFT22 specifically associates with G-nucleotides and present crystal structures of IFT22 in complex with GDP, GTP, and with IFT74/81. Our structural analysis unravels an unusual GTP/GDP-binding mode of IFT22 bypassing the classical G4 motif. The GTPase switch regions of IFT22 become ordered upon complex formation with IFT74/81 and mediate most of the IFT22-74/81 interactions. Structure-based mutagenesis reveals that association of IFT22 with the IFT complex is essential for flagellum construction in Trypanosoma brucei although IFT22 GTP-loading is not strictly required.

KW - cilia

KW - GTPase

KW - IFT22

KW - intraflagellar transport

KW - Trypanosoma brucei

UR - http://www.scopus.com/inward/record.url?scp=85063799499&partnerID=8YFLogxK

U2 - 10.15252/embj.2018101251

DO - 10.15252/embj.2018101251

M3 - Journal article

C2 - 30940671

AN - SCOPUS:85063799499

SN - 0261-4189

VL - 38

JO - EMBO Journal

JF - EMBO Journal

IS - 9

M1 - e101251

ER -

Binding of IFT22 to the intraflagellar transport complex is essential for flagellum assembly

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Keywords

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