TIAM-1/GEF can shape somatosensory dendrites independently of its GEF activity by regulating F-actin localization
- Albert Einstein College of Medicine, United States
Figures
Figure 1 with 1 supplement
The intracellular domain of DMA-1/LRR-TM is required for higher order branching of PVD somatosensory dendrites.
(A) Fluorescent images of PVD (left panels) and schematics (right panels) of wild-type control animals. PVD is visualized by the wdIs52 [F49H12.4p::GFP] transgene in all panels. 1°, 2°, 3°, 4°, and …
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Figure 1—source data 1
Complete source data.
- https://doi.org/10.7554/eLife.38949.003
Figure 1—figure supplement 1
Genes functioning cell-autonomously in PVD somatosensory neurons.
(A–B). Genomic environs of the indicated genes with the physical location on the respective linkage groups (LGs) are shown. The exon-intron structure is indicated, as is the direction of …
Figure 2 with 2 supplements
The hpo-30/Claudin, tiam-1/GEF, and act-4/Actin act genetically in the Menorin pathway.
(A – D) Fluorescent images of PVD (left panels, visualized by the wdIs52 [F49H12.4p::GFP] transgene) and schematics (right panels) of the genotypes indicated. The control image is identical to Figure…
Figure 2—figure supplement 1
Effects of hpo-30/Claudin, tiam-1/GEF and act-4/Actin on localization of LECT-2:mNG, SAX-7::GFP and DMA-1::GFP.
(A) Fluorescent images of animals expressing GFP in FLP neurons (muIs32 [Pmec-7::GFP]) in the indicated genotypes. Anterior is to the left and scale bars indicate 20 µm. (B – D) Fluorescent images …
Figure 2—figure supplement 2
The Genetics of hpo-30/Claudin, tiam-1/GEF, and act-4/Actin.
(A – H) Fluorescent images of animals expressing GFP in PVD neurons (wdIs52 [F49H12.4p::GFP]) in the indicated genotypes. Anterior is to the left and scale bars indicate 20 µm. (I – K) …
Figure 3 with 1 supplement
The tiam-1/GEF and act-4/Actin act downstream of the dma-1/LRR-TM receptor in PVD dendrites.
(A – D) Fluorescent images of animals in which mnr-1/Menorin was expressed ectopically in muscle (mnr-1(gof): dzIs43, [myo-3p::mnr-1] (Salzberg et al., 2013)) in different genetic backgrounds. A …
Figure 3—figure supplement 1
The tiam-1/GEF and act-4/Actin act downstream or in parallel of the dma-1/LRR-TM receptor in PVD dendrites.
Quantification of the aggregate length of secondary, tertiary, and quaternary branches 100 µm anterior to the PVD cell body in the genotypes indicated. For raw data see Figure 1— source data 1. …
Figure 4
HPO-30/Claudin forms a complex with, and, localizes DMA-1/LRR-TM to higher order branches of PVD somatosensory neurons.
(A – D) Images of an animal with PVD-specific expression of HPO-30::tagBFP (A, dzEx1837) and DMA-1::GFP (B, qyIs369) as well as a composite image (C), respectively. Both reporters show diffuse …
Figure 5 with 2 supplements
TIAM-1/GEF interacts with DMA-1/LRR-TM through PDZ motif/domain interaction to shape 4° dendrite branches.
(A – B) Fluorescent image with schematics of tiam-1 mutant animals carrying either a full length or deletion construct (GEF-only) of the tiam-1 cDNA under control of the PVD-specific ser-2p3 …
Figure 5—figure supplement 1
The TIAM-1/GEF acts independently of Rac1 GEF activity.
images of animals expressing GFP in PVD neurons (wdIs52 [F49H12.4p::GFP]) in the indicated genotypes. Anterior is to the left in A and to the right in B and C. (D) Quantification of animals with a …
Figure 5—figure supplement 2
Supplementary co-immunoprecipitation experiments (related to Figure 5 (A) Figure 7 (B)).
(A) Replacement of PDZ motif in DMA-1 with EFYA (SDN-1 PDZ motif) diminishes the binding affinity between TIAM-1 and DMA-1. Western blots of co-immunoprecipitation experiments. Transfected …
Figure 6
The TIAM-1/GEF functions independently of its guanine nucleotide activity.
(A) Genomic environs of the tiam-1 locus in linkage group I (LGI) with exons and introns indicated. The location of the two CRISPR/Cas9 engineered dz264 and dz265 alleles, encoding a missense …
Figure 7 with 1 supplement
F-actin is localized to the leading edges of developing dendrites and requires the Menorin pathway for polarized localization.
(A) Fluorescent images of animals at different developmental stages carrying a F-actin reporter (ser-2p3::tagRFP::UtrCH [dzEx1564], left panels), a PVD cytoplasmic GFP reporter (wdIs52 …
Figure 7—figure supplement 1
Localization of TBA-1::tagRFP in different genetic backgrounds.
Fluorescent images of animals in different genetic backgrounds carrying a microtubule reporter (dzEx1569, Ex[ser-2p3::TBA-1::tagRFP], left panels), a PVD cytoplasmic GFP reporter (middle panels), …
Additional files
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Supplementary file 1
Lists of all mutants alleles used, as well as all plasmids with details of their construction.
In addition, Supplementary file 1 lists all transgenic strains created during the course of these studies.
- https://doi.org/10.7554/eLife.38949.023
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Transparent reporting form
- https://doi.org/10.7554/eLife.38949.024
