Fig. 4

KIAA0556 is a basal body and MT-associated protein. a GFP- and Strep/FLAG tagged KIAA0556 is localised to the basal body of the primary cilium of RPE1 cells. Cells were counterstained for RPGRIP1L (cyan; transition zone marker) and polyglutamylated tubulin (PolyGluTub; red; ciliary and basal body marker). Scale bars, 20 μm. b A C. elegans KIAA0556 (K04F10.2) transcriptional reporter is expressed almost exclusively in ciliated cells (amphids, PHA/B phasmids, PDE, labial/quadrant neurons). Expression is also detected in two intestinal cells, Int1/Int8. Scale bars, 20 μm (whole worm), 5 μm (insets). c, d C. elegans K04F10.2::GFP accumulates at the ciliary base, proximal to the transition zone, and within the proximal ciliary axoneme (middle segment and transition zone). Schematic in (c) denotes the MT architecture of amphid and phasmid channel cilia, consisting of a distal segment (ds; singlet A tubules), middle segment (ms; doublet A/B tubules), transition zone (tz; containing Y-links) and proximal MT flares (analogous to a basal body; bb) in the periciliary membrane compartment (pcmc). The red arrows denote that the isosceles trapezoid shape of the K04F10.2::GFP ciliary base signal corresponds to that of the transmission electron microscopy-defined MT flares [29]. The kymograph (time over distance plot; t/d) in (c) is derived from a time lapse video and indicates that K04F10.2::GFP does not undergo processive movement in cilia. White arrows in (c) denote ciliary base accumulation of GFP signal. Fluorescence images in (d) captured from worms expressing K04F10.2::GFP and MKS-5::tdTomato transgenes show that a major pool of K04F10.2::GFP signal lies immediately proximal to TZ-localised MKS-5. CB cell body. Scale bars, 3 μm