1 Genes
| WormBase Gene ID | Gene Name | Sequence Name | Organism |
|---|---|---|---|
| WBGene00001248 | elp-1 | F38A6.2 | Caenorhabditis elegans |
| Primary Identifier | Expr8966 | Remark | Picture: Fig 6, 7, 8, 9. |
| Subcellular Localization | Affinity-purified antibodies against ELP-1 and the full-length ELP-1::GFP fusion protein localized to thin linear rows near the cell surface that were not quite parallel to the long axis of the muscle cell. At higher resolution there was a periodicity to these lines that approximated the distance between the dense body adhesion sites. The ELP-1::GFP fluorescence slightly overlaps the edges of the dense bodies but is not superimposable with the dense bodies at this angle. These results indicate that ELP-1 is unlikely to be a component of the dense bodies. Double-stained muscle cells with anti-ELP-1 antibodies and a monoclonal antibody MH25 against PAT-3 shows that integrin and ELP-1 have overlapping staining patterns at this level of resolution. Because of the relatively impenetrable cuticle that surrounds the nematode, these worms were frozen and cracked open prior to antibody staining. In areas lacking dense bodies, ELP-1::GFP was sometimes lost. However most of the ELP-1::GFP was retained in a linear pattern. These results indicate that the membrane and dense bodies can be separated from the ELP-1::GFP. Although the truncated construct and the full-length construct were expressed in the same cells and tissues, only the full length ELP-1::GFP construct localized to an elaborate array of fluorescent filaments. An obliquely striated fluorescent pattern emerges approximately 0.8 um apically from the base of the dense body. Deeper within the muscle body the filaments appear to separate from the linear track and branch out into the cytoplasm. ELP-1 was enriched in preparations of paclitaxel-stabilized microtubules in vitro. Paclitaxel-stabilized microtubules were prepared from a mixed-stage worm preparation and examined by means of Western blotting with an affinity-purified antisera against a bacterially expressed ELP-1 fusion protein. An ELP-1-reacting band co-purified with microtubules and migrated at ~100 kDa, a mass that corresponded to the Genefinder predictions for the larger ELP-1a polypeptide. Two smaller and less abundant proteins that cross-react with the ELP-1 antibodies also co-purified with microtubules. These may be isoforms generated by alternative splicing (i.e. ELP-1b) or proteolytic fragments of ELP-1a. This experiment and the one described above showed that ELP-1 is microtubule-associated both in situ and in vivo. To test the idea that the filaments decorated by ELP-1::GFP were microtubules, authors treated worms with the microtubule inhibitor, nocodazole. Worms were gently pressed between a slide and coverslip to extrude their intestines. This process was done in the presence of levamisole, an acetylcholine agonist, which was used to paralyze the worms for microscopy. The filaments in the extruded layer were stable for 30 minutes or more in the absence of nocodazole. The decorated filaments began to degrade within seconds and were entirely de-polymerized after two minutes in nocodazole. When the fluorescence disappeared the intestinal fragments noticeably flattened and the remnants of the worm contracted. This result indicates that ELP-1::GFP associates with microtubules in situ. |
| WormBase Gene ID | Gene Name | Sequence Name | Organism |
|---|---|---|---|
| WBGene00001248 | elp-1 | F38A6.2 | Caenorhabditis elegans |
