Sam Li / Jose-Jesus Fernandez / Amy S Fabritius / David A Agard / Mark Winey /
PubMed Abstract
Doublet microtubules (DMTs) provide a scaffold for axoneme assembly in motile cilia. Aside from α/β tubulins, the DMT comprises a large number of non-tubulin proteins in the luminal wall of DMTs, ...Doublet microtubules (DMTs) provide a scaffold for axoneme assembly in motile cilia. Aside from α/β tubulins, the DMT comprises a large number of non-tubulin proteins in the luminal wall of DMTs, collectively named the microtubule inner proteins (MIPs). We used cryoET to study axoneme DMT isolated from We present the structures of DMT at nanometer and sub-nanometer resolution. The structures confirm that MIP RIB72A/B binds to the luminal wall of DMT by multiple DM10 domains. We found FAP115, an MIP-containing multiple EF-hand domains, located at the interface of four-tubulin dimers in the lumen of A-tubule. It contacts both lateral and longitudinal tubulin interfaces and playing a critical role in DMT stability. We observed substantial structure heterogeneity in DMT in an knockout strain, showing extensive structural defects beyond the FAP115-binding site. The defects propagate along the axoneme. Finally, by comparing DMT structures from and , we have identified a number of conserved MIPs as well as MIPs that are unique to each organism. This conservation and diversity of the DMT structures might be linked to their specific functions. Our work provides structural insights essential for understanding the roles of MIPs during motile cilium assembly and function, as well as their relationships to human ciliopathies.
EMDB-24364: A-tubule of axonemal microtubule doublet in 16 nm repeat from Tetrahymena thermophila Method: EM (subtomogram averaging) / Resolution: 10.2 Å
EMDB-24366: B-tubule of axonemal microtubule doublet in 16 nm repeat from Tetrahymena thermophila Method: EM (subtomogram averaging) / Resolution: 10.2 Å
EMDB-24367: Partial of A-tubule of axonemal microtubule doublet in 16 nm repeat from Tetrahymena thermophila Method: EM (subtomogram averaging) / Resolution: 9.9 Å
EMDB-24368: A-tubule of axonemal microtubule doublet in 48 nm repeat from Tetrahymena thermophila Method: EM (subtomogram averaging) / Resolution: 12.0 Å
EMDB-24370: B-tubule of axonemal microtubule doublet in 48 nm repeat from Tetrahymena thermophila Method: EM (subtomogram averaging) / Resolution: 12.0 Å
EMDB-24371: A-tubule of axonemal microtubule doublet in 16 nm repeat from Tetrahymena thermophila mutant fap115 knockout Method: EM (subtomogram averaging) / Resolution: 9.6 Å
EMDB-24372: B-tubule of axonemal microtubule doublet in 16 nm repeat from Tetrahymena thermophila mutant fap115 knockout Method: EM (subtomogram averaging) / Resolution: 9.6 Å
EMDB-24373: Partial A-tubule of axonemal microtubule doublet in 16 nm repeat from Tetrahymena thermophila mutant fap115 knockout Method: EM (subtomogram averaging) / Resolution: 9.6 Å
EMDB-24374: A-tubule of axonemal microtubule doublet in 48 nm repeat from Tetrahymena thermophila mutant fap115 knockout Method: EM (subtomogram averaging) / Resolution: 11.8 Å
EMDB-24375: B-tubule of axonemal microtubule doublet in 48 nm repeat from Tetrahymena thermophila mutant fap115 knockout Method: EM (subtomogram averaging) / Resolution: 12.0 Å
EMDB-24376: Inner junction structure of axonemal microtubule doublet in 48 nm repeat from Tetrahymena thermophila Method: EM (subtomogram averaging) / Resolution: 12.0 Å
EMDB-24377: An unidentified MIP at the inner junction of axonemal microtubule doublet from Tetrahymena thermophila Method: EM (subtomogram averaging) / Resolution: 10.9 Å
EMDB-24379: Composite map of axonemal microtubule doublet in 16 nm repeat from Tetrahymena thermophila Method: EM (subtomogram averaging) / Resolution: 10.2 Å
EMDB-24380: Composite map of axonemal microtubule doublet in 48 nm repeat from Tetrahymena thermophila Method: EM (subtomogram averaging) / Resolution: 12.0 Å
EMDB-24381: Composite map of axonemal microtubule doublet in 16 nm repeat from a Tetrahymena thermophila mutant fap115 knockout Method: EM (subtomogram averaging) / Resolution: 9.6 Å
EMDB-24382: Composite map of axonemal microtubule doublet in 48 nm repeat from a Tetrahymena thermophila mutant fap115 knockout Method: EM (subtomogram averaging) / Resolution: 12.0 Å
Source
Tetrahymena thermophila (eukaryote)
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Tetrahymena thermophila (eukaryote)