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2015 ; 35
(4
): 497-512
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Proteomics of Primary Cilia by Proximity Labeling
#MMPMID26585297
Mick DU
; Rodrigues RB
; Leib RD
; Adams CM
; Chien AS
; Gygi SP
; Nachury MV
Dev Cell
2015[Nov]; 35
(4
): 497-512
PMID26585297
show ga
While cilia are recognized as important signaling organelles, the extent of
ciliary functions remains unknown because of difficulties in cataloguing proteins
from mammalian primary cilia. We present a method that readily captures rapid
snapshots of the ciliary proteome by selectively biotinylating ciliary proteins
using a cilia-targeted proximity labeling enzyme (cilia-APEX). Besides
identifying known ciliary proteins, cilia-APEX uncovered several ciliary
signaling molecules. The kinases PKA, AMPK, and LKB1 were validated as bona fide
ciliary proteins and PKA was found to regulate Hedgehog signaling in primary
cilia. Furthermore, proteomics profiling of Ift27/Bbs19 mutant cilia correctly
detected BBSome accumulation inside Ift27(-/-) cilia and revealed that ?-arrestin
2 and the viral receptor CAR are candidate cargoes of the BBSome. This work
demonstrates that proximity labeling can be applied to proteomics of
non-membrane-enclosed organelles and suggests that proteomics profiling of cilia
will enable a rapid and powerful characterization of ciliopathies.
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