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. 2019 Jan 29;8:e38949. doi: 10.7554/eLife.38949

Figure 6. The TIAM-1/GEF functions independently of its guanine nucleotide activity.

Figure 6.

(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 mutation that causes the T548F mutation is shown. (B) Fluorescent images with schematics of tiam-1 mutant animals carrying a T548F point mutant form of the tiam-1 cDNA under control of the PVD-specific ser-2p3 promoter. The T548F point mutant is analogous to the S1216F mutation in UNC-73 (Steven et al., 1998) and the T516F mutation in a previously described TIAM-1 cDNA (Demarco et al., 2012). Importantly, the point mutations in both proteins have been shown to abrogate GEF activity toward Rac in vitro (Demarco et al., 2012). (C – D) Fluorescent images of PVD (left panels) and schematics (right panels) of wild-type control (C) and tiam-1(dz265) mutant animals (D). PVD is visualized by the wdIs52 [F49H12.4p::GFP] transgene in all panels.