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J Cell Sci. 2017 Mar 15;130(6):1110-1121. doi: 10.1242/jcs.200006. Epub 2017 Feb 2.

The T cell IFT20 interactome reveals new players in immune synapse assembly.

Author information

1
Department of Life Sciences, University of Siena, Siena 53100, Italy.
2
Sir William Dunn School of Pathology, University of Oxford, Oxford OX1 3RE, UK.
3
Department of Biotechnology, Chemistry and Pharmacy, University of Siena, Siena 53100, Italy.
4
Department of Life Sciences, University of Siena, Siena 53100, Italy cosima.baldari@unisi.it.

Abstract

Sustained signalling at the immune synapse (IS) requires the synaptic delivery of recycling endosome-associated T cell antigen receptors (TCRs). IFT20, a component of the intraflagellar transport system, controls TCR recycling to the IS as a complex with IFT57 and IFT88. Here, we used quantitative mass spectrometry to identify additional interaction partners of IFT20 in Jurkat T cells. In addition to IFT57 and IFT88, the analysis revealed new binding partners, including IFT54 (also known as TRAF3IP1), GMAP-210 (also known as TRIP11), Arp2/3 complex subunit-3 (ARPC3), COP9 signalosome subunit-1 (CSN1, also known as GPS1) and ERGIC-53 (also known as LMAN1). A direct interaction between IFT20 and both IFT54 and GMAP-210 was confirmed in pulldown assays. Confocal imaging of antigen-specific conjugates using T cells depleted of these proteins by RNA interference showed that TCR accumulation and phosphotyrosine signalling at the IS were impaired in the absence of IFT54, ARPC3 or ERGIC-53. Similar to in IFT20-deficient T cells, this defect resulted from a reduced ability of endosomal TCRs to polarize to the IS despite a correct translocation of the centrosome towards the antigen-presenting cell contact. Our data underscore the traffic-related role of an IFT20 complex that includes components of the intracellular trafficking machinery in IS assembly.

KEYWORDS:

Immune synapse assembly; Intraflagellar transport system; Mass spectrometry analysis

PMID:
28154159
PMCID:
PMC5358343
DOI:
10.1242/jcs.200006
[Indexed for MEDLINE]
Free PMC Article

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