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TitleTetrahymena RIB72A and RIB72B are microtubule inner proteins in the ciliary doublet microtubules.
Journal, issue, pagesMol Biol Cell, Vol. 29, Issue 21, Page 2566-2577, Year 2018
Publish dateOct 15, 2018
AuthorsDaniel Stoddard / Ying Zhao / Brian A Bayless / Long Gui / Panagiota Louka / Drashti Dave / Swati Suryawanshi / Raphaël F-X Tomasi / Pascale Dupuis-Williams / Charles N Baroud / Jacek Gaertig / Mark Winey / Daniela Nicastro /
PubMed AbstractDoublet and triplet microtubules are essential and highly stable core structures of centrioles, basal bodies, cilia, and flagella. In contrast to dynamic cytoplasmic micro-tubules, their luminal ...Doublet and triplet microtubules are essential and highly stable core structures of centrioles, basal bodies, cilia, and flagella. In contrast to dynamic cytoplasmic micro-tubules, their luminal surface is coated with regularly arranged microtubule inner proteins (MIPs). However, the protein composition and biological function(s) of MIPs remain poorly understood. Using genetic, biochemical, and imaging techniques, we identified Tetrahymena RIB72A and RIB72B proteins as ciliary MIPs. Fluorescence imaging of tagged RIB72A and RIB72B showed that both proteins colocalize to Tetrahymena cilia and basal bodies but assemble independently. Cryoelectron tomography of RIB72A and/or RIB72B knockout strains revealed major structural defects in the ciliary A-tubule involving MIP1, MIP4, and MIP6 structures. The defects of individual mutants were complementary in the double mutant. All mutants had reduced swimming speed and ciliary beat frequencies, and high-speed video imaging revealed abnormal highly curved cilia during power stroke. Our results show that RIB72A and RIB72B are crucial for the structural assembly of ciliary A-tubule MIPs and are important for proper ciliary motility.
External linksMol Biol Cell / PubMed:30133348 / PubMed Central
MethodsEM (subtomogram averaging)
Resolution40.1 Å
Structure data

EMDB-7805:
Subtomogram average (699 axonemal repeats) of isolated Rib72A/B double knock out Tetrahymena cilia showing the lumen of the ciliary doublet microtubule (DMT)
Method: EM (subtomogram averaging) / Resolution: 40.1 Å

EMDB-7806:
Subtomogram average (758 axonemal repeats) of isolated Rib72A knock out Tetrahymena cilia showing the lumen of the ciliary doublet microtubule (DMT)
Method: EM (subtomogram averaging) / Resolution: 40.1 Å

EMDB-7807:
Subtomogram average (735 axonemal repeats) of isolated Rib72B knock out Tetrahymena cilia Rescued with Rib72B-GFP, showing the lumen of the ciliary doublet microtubule (DMT)
Method: EM (subtomogram averaging) / Resolution: 40.1 Å

EMDB-7811:
Subtomogram average (702 axonemal repeats) of isolated Rib72B knock out Tetrahymena cilia showing the lumen of the ciliary doublet microtubule (DMT)
Method: EM (subtomogram averaging) / Resolution: 40.1 Å

Source
  • Tetrahymena thermophila (eukaryote)

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