Axoneme-specific β-tubulin specialization: a conserved C-terminal motif specifies the central pair

Abstract

Axonemes are ancient organelles that mediate motility of cilia and flagella in animals, plants, and protists. The long evolutionary conservation of axoneme architecture, a cylinder of nine doublet microtubules surrounding a central pair of singlet microtubules, suggests all motile axonemes may share common assembly mechanisms. Consistent with this, α- and β-tubulins utilized in motile axonemes fall among the most conserved tubulin sequences [1, 2], and the β-tubulins contain a sequence motif at the same position in the carboxyl terminus [3]. Axoneme doublet microtubules are initiated from the corresponding triplet microtubules of the basal body [4], but the large macromolecular “central apparatus” that includes the central pair microtubules and associated structures [5] is a specialization unique to motile axonemes. In Drosophila spermatogenesis, basal bodies and axonemes utilize the same α-tubulin but different β-tubulins [6–13]. β1 is utilized for the centriole/basal body, and β2 is utilized for the motile sperm tail axoneme. β2 contains the motile axoneme-specific sequence motif, but β1 does not [3]. Here, we show that the “axoneme motif” specifies the central pair. β1 can provide partial function for axoneme assembly but cannot make the central microtubules [14]. Introducing the axoneme motif into the β1 carboxyl terminus, a two amino acid change, conferred upon β1 the ability to assemble 9 + 2 axonemes. This finding explains the conservation of the axoneme-specific sequence motif through 1.5 billion years of evolution.