The Caenorhabditis elegans Tubby homolog dynamically modulates olfactory cilia membrane morphogenesis and phospholipid composition
- Brandeis University, United States
Figures
Figure 1 with 1 supplement
TUB-1 is necessary for membrane morphogenesis in wing cilia.
(A) Representative images of AWA, AWC and AWB cilia in wild-type and tub-1(nr2004) mutants. Cartoons of cilia morphologies are shown at top. Numbers at top right (in AWA and AWC image panels) …
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Figure 1—source data 1
Data for Figure 1B and E, and Figure 1—figure supplement 1.
- https://doi.org/10.7554/eLife.48789.004
Figure 1—figure supplement 1
TUB-1 regulates AWB cilia morphology.
Representative images (top) and quantification (bottom) of AWB cilia lengths in animals of the indicated genotypes. Human (Hs - Homo sapiens) GFP::TULP1 and GFP::TULP3-encoding sequences were …
Figure 2 with 1 supplement
TUB-1 regulates ciliary targeting of transmembrane proteins in AWB.
(A–G) Representative images of localization patterns of the indicated fusion proteins in wild-type (WT) and tub-1(nr2004) mutants in AWB (A–E), ASK (F) and ASH (G). The AWB, ASK, and ASH neurons …
Figure 2—figure supplement 1
TAX-4 localization is unaltered in odr-3(gof) mutants.
Representative images of str-1p::mCherry and str-1p::TAX-4::GFP in AWB neurons of wild-type and odr-3(Q206L)XS (Roayaie et al., 1998) animals. The AWB neuron is marked via expression of str-1p::mCher…
Figure 3 with 1 supplement
TUB-1 acts both at the AWB PCMC and within cilia to regulate AWB cilia morphology.
(A) Representative images of tagRFP::TUB-1 localization in AWB indicating cytoplasmic localization in the soma (open orange arrow) and enrichment at the dendritic ends (yellow arrowheads; left and …
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Figure 3—source data 1
Data for Figure 3D,E,F, and Figure 3—figure supplement 1A.
- https://doi.org/10.7554/eLife.48789.009
Figure 3—figure supplement 1
tub-1 shares a subset of phenotypes with daf-10.
(A) Representative images (left) and quantification of AWB cilia lengths (right) in animals of the indicated genotypes. Alleles used were tub-1(nr2004) and daf-10(e1387). *** indicates different …
Figure 4 with 1 supplement
Reduced sensory signaling increases TUB-1 localization within AWB cilia.
(A) (Top) Representative images of AWB cilia morphologies in animals of the indicated genotypes. Alleles used were tub-1(nr2004) and odr-1(n1936). The AWB neurons were visualized via expression of st…
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Figure 4—source data 1
Data for Figure 4A,B,D,E, and Figure 4—figure supplement 1.
- https://doi.org/10.7554/eLife.48789.012
Figure 4—figure supplement 1
Loss-of-function mutations in daf-10 suppress the expanded ‘fan’ phenotype of odr-1 mutants.
Representative images (top) and quantification of maximum AWB cilia widths in animals of the indicated genotypes. *** and &&& indicate different from wild-type and odr-1, respectively, at p<0.001 …
Figure 5 with 1 supplement
Levels of PI(4,5)P2 and PPK-1 are increased within AWB cilia in odr-1 mutants.
(A, C, D) Representative images of GFP::PLCδ1-PH (A), tagRFP::TUB-1 (C) and GFP::PPK-1 (D) localization in AWB cilia of animals of the indicated genotypes. Alleles used were odr-1(n1936) and inpp-1(g…
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Figure 5—source data 1
Data for Figures 5B and E, and Figure 5—figure supplement 1.
- https://doi.org/10.7554/eLife.48789.015
Figure 5—figure supplement 1
INPP-1 localization is unaltered in tub-1 and odr-1 mutants.
(A) Representative images of INPP-1A::GFP localization in AWB cilia of animals of the indicated genotypes. Yellow and white arrowheads indicate the cilia base and cilia, respectively; arrow …
Figure 6 with 2 supplements
TUB-1 regulates PPK-1 localization and PI(4,5)P2 distribution in AWB dendrites and cilia.
(A, D) Representative images of GFP::PPK-1 (A) and GFP::PLCδ1-PH (D) localization in animals of the indicated genotypes. Alleles used were tub-1(nr2004) and odr-1(n1936). The AWB neuron is marked …
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Figure 6—source data 1
Data for Figure 6B,C,E,F, Figure 6—figure supplement 1, and Figure 6—figure supplement 2B and C.
- https://doi.org/10.7554/eLife.48789.019
Figure 6—figure supplement 1
TUB-1 may act cell-autonomously to regulate GFP::PLCδ1-PH distribution in AWB.
Representative images (left) and line scans (right) of GFP::PLCδ1-PH distribution in AWB cilia and dendrites in animals of the indicated genotypes. Yellow and white arrowheads indicate the cilia …
Figure 6—figure supplement 2
TUB-1 does not regulate PPK-1 and PI(4,5)P2 distribution in ASK dendrites and cilia.
(A) Representative images of GFP::TUB-1 localization in the ASK neurons and enrichment at the dendritic ends (yellow arrowheads). ASK soma is indicated by an open orange arrowhead. GFP::TUB-1 is …
Figure 7 with 1 supplement
TUB-1 acts in part via the DPY-23 AP-2 μ subunit to localize PPK-1 at the PCMC.
(A, B) (Left) Representative images of DPY-23::GFP (A) and GFP::PLCδ1-PH (B) localization in AWB cilia of animals of the indicated genotypes. AWB cilia were visualized via expression of str-1p::mCher…
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Figure 7—source data 1
Data for Figure 7A–D and Figure 7—figure supplement 1A–C.
- https://doi.org/10.7554/eLife.48789.022
Figure 7—figure supplement 1
Localization patterns of endocytic proteins in AWB and ASK in tub-1 mutants.
(A–C) Representative images (left) and quantification of fluorescence intensities (right) of the indicated proteins at the PCMCs of AWB (A) and ASK (B and C). (Left images) Neurons are marked via …
Figure 8
Model for TUB-1 function in regulating AWB olfactory cilia properties.
Cartoons of proposed TUB-1 functions at the PCMC and within AWB cilia. Dotted yellow bars indicate singlet microtubules present in the distal segments of AWB cilia; the origins of these singlet …
Additional files
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Supplementary file 1
List of strains used in this work.
- https://doi.org/10.7554/eLife.48789.024
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Supplementary file 2
List of plasmids used in this work.
- https://doi.org/10.7554/eLife.48789.025
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Transparent reporting form
- https://doi.org/10.7554/eLife.48789.026
