Type of Submission
Poster
Keywords
Tetrahymena, netrin, tyrosine kinase, UNC-5
Proposal
Netrins are pleiotropic signaling molecules with diverse roles in animal development. Netrin signals through a number of receptors in animals, including the UNC-5 family, neogenin, DSCAM, and DCC. Previous studies have shown that netrin-1-peptide, netrin-3-peptide, and recombinant netrin-4 all act as chemorepellents in Tetrahymena (Kuruvilla et al., 2016, Khol et al., 2018; Bradley and Kuruvilla, 2020). In addition, netrin-1 peptide appears to signal through a tyrosine kinase in this organism (Kuruvilla et al., 2016), similar to vertebrate signaling through UNC-5, which uses the tyrosine kinase, src. In light of these data, we hypothesized that Tetrahymena thermophila possess netrin signaling machinery, including a tyrosine kinase. In order to investigate this hypothesis, we searched for various netrin receptors, as well as a src homologue, in Tetrahymena using immunofluorescence (Khol et al., 2018). We found that anti-UNC-5 and anti-neogenin antibodies showed fluorescence, while anti-DCC and anti-DSCAM antibodies did not. In addition, an anti-src antibody showed significant fluorescence in Tetrahymena (Khol et al., 2018. In our current study, we searched the Tetrahymena Genome Database for homologs of UNC-5, neogenin, and src. We also used Western blotting to screen for potential homologues of these proteins. At the present time, there are several proteins of interest which we would like to study further.
Start Date
4-8-2020 1:00 PM
End Date
4-22-2020 6:00 PM
Creative Commons License
This work is licensed under a Creative Commons Attribution-Noncommercial-No Derivative Works 4.0 License.
How the Western Was Won: Evidence for Netrin Signaling Machinery in Tetrahymena thermophila
Netrins are pleiotropic signaling molecules with diverse roles in animal development. Netrin signals through a number of receptors in animals, including the UNC-5 family, neogenin, DSCAM, and DCC. Previous studies have shown that netrin-1-peptide, netrin-3-peptide, and recombinant netrin-4 all act as chemorepellents in Tetrahymena (Kuruvilla et al., 2016, Khol et al., 2018; Bradley and Kuruvilla, 2020). In addition, netrin-1 peptide appears to signal through a tyrosine kinase in this organism (Kuruvilla et al., 2016), similar to vertebrate signaling through UNC-5, which uses the tyrosine kinase, src. In light of these data, we hypothesized that Tetrahymena thermophila possess netrin signaling machinery, including a tyrosine kinase. In order to investigate this hypothesis, we searched for various netrin receptors, as well as a src homologue, in Tetrahymena using immunofluorescence (Khol et al., 2018). We found that anti-UNC-5 and anti-neogenin antibodies showed fluorescence, while anti-DCC and anti-DSCAM antibodies did not. In addition, an anti-src antibody showed significant fluorescence in Tetrahymena (Khol et al., 2018. In our current study, we searched the Tetrahymena Genome Database for homologs of UNC-5, neogenin, and src. We also used Western blotting to screen for potential homologues of these proteins. At the present time, there are several proteins of interest which we would like to study further.
