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The C‐terminal tails of heterotrimeric kinesin‐2 motor subunits directly bind to α‐tubulin1: Possible implications for cilia‐specific tubulin entry
Author(s) -
Girotra Mukul,
Srivastava Shalini,
Kulkarni Anuttama,
Barbora Ayan,
Bobra Kratika,
Ghosal Debnath,
Devan Pavithra,
Aher Amol,
Jain Akanksha,
Panda Dulal,
Ray Krishanu
Publication year - 2017
Publication title -
traffic
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 2.677
H-Index - 130
eISSN - 1600-0854
pISSN - 1398-9219
DOI - 10.1111/tra.12461
Subject(s) - kinesin , microtubule , intraflagellar transport , cilium , biology , microbiology and biotechnology , tubulin , heterotrimeric g protein , cytoskeleton , motor protein , fluorescence recovery after photobleaching , flagellum , biochemistry , g protein , signal transduction , gene , cell , membrane
The assembly of microtubule‐based cytoskeleton propels the cilia and flagella growth. Previous studies have indicated that the kinesin‐2 family motors transport tubulin into the cilia through intraflagellar transport. Here, we report a direct interaction between the C‐terminal tail fragments of heterotrimeric kinesin‐2 and α‐tubulin1 isoforms in vitro. Blot overlay screen, affinity purification from tissue extracts, cosedimentation with subtilisin‐treated microtubule and LC‐ESI‐MS / MS characterization of the tail‐fragment‐associated tubulin identified an association between the tail domains and α‐ tubulin1A /D isotype. The interaction was confirmed by Forster's resonance energy transfer assay in tissue‐cultured cells. The overexpression of the recombinant tails in NIH3T3 cells affected the primary cilia growth, which was rescued by coexpression of a α‐tubulin1 transgene. Furthermore, fluorescent recovery after photobleach analysis in the olfactory cilia of Drosophila indicated that tubulin is transported in a non‐particulate form requiring kinesin‐2. These results provide additional new insight into the mechanisms underlying selective tubulin isoform enrichment in the cilia.