Abstract
Cilia and flagella, which play essential roles in cell motility, sensing and development, contain at their core a microtubule-based structure called axoneme. The axoneme, which contains nine doublet and two central microtubules, is highly stable and its length is precisely-controlled. We have asked whether the axonemal tubulins contribute to this length stability. Towards this end, we used a novel procedure to differentially extract the tubulins from the different axoneme components, and characterized their dynamic properties and post-translational modifications (PTMs). We found that their dynamic properties are consistent with the greater stability of axonemes. Despite the differences in PTMs between the axonemal components, we found no significant differences in their dynamic properties. Unexpectedly, the reconstituted ciliary microtubules exhibit curved protofilaments at their growing tip, that may correspond to the fluctuating GTP-cap hypothesized to exist. Thus, our study provides new insights into the growth, stability and the role of PTMs of ciliary tubulin.
Abbreviations
- MTs
- microtubules;
- PTMs
- post-translational modifications;
- PFs
- protofilaments;
- CP
- central pair;
- MAPs
- microtubule-associated proteins;
- IRM
- interference reflection microscopy;
- TEM
- transmission electron microscopy;
- cryo-EM
- cryo-electron microscopy;
- MSD
- mean squared displacement;
- GMP-CPP
- guanosine-5’-[(α,β)methyleno]triphosphate
Abbreviations
- MTs
- microtubules;
- PTMs
- post-translational modifications;
- PFs
- protofilaments;
- CP
- central pair;
- MAPs
- microtubule-associated proteins;
- IRM
- interference reflection microscopy;
- TEM
- transmission electron microscopy;
- cryo-EM
- cryo-electron microscopy;
- MSD
- mean squared displacement;
- GMP-CPP
- guanosine-5’-[(α,β)methyleno]triphosphate