Welcome to the new version of CaltechAUTHORS. Login is currently restricted to library staff. If you notice any issues, please email coda@library.caltech.edu
Published June 2009 | Supplemental Material + Published
Journal Article Open

The cell surface receptor Tartan is a potential in vivo substrate for the receptor tyrosine phosphatase Ptp52F


Receptor-linked protein-tyrosine phosphatases (RPTPs) are essential regulators of axon guidance and synaptogenesis in Drosophila, but the signaling pathways in which they function are poorly defined. We identified the cell surface receptor Tartan (Trn) as a candidate substrate for the neuronal RPTP Ptp52F by using a modified two-hybrid screen with a substrate-trapping mutant of Ptp52F as "bait." Trn can bind to the Ptp52F substrate-trapping mutant in transfected Drosophila S2 cells if v-Src kinase, which phosphorylates Trn, is also expressed. Coexpression of wild-type Ptp52F causes dephosphorylation of v-Src-phosphorylated Trn. To examine the specificity of the interaction in vitro, we incubated Ptp52F-glutathione S-transferase (GST) fusion proteins with pervanadate-treated S2 cell lysates. Wild-type Ptp52F dephosphorylated Trn, as well as most other bands in the lysate. GST "pulldown" experiments demonstrated that the Ptp52F substrate-trapping mutant binds exclusively to phospho-Trn. Wild-type Ptp52F pulled down dephosphorylated Trn, suggesting that it forms a stable Ptp52F-Trn complex that persists after substrate dephosphorylation. To evaluate whether Trn and Ptp52F are part of the same pathway in vivo, we examined motor axon guidance in mutant embryos. trn and Ptp52F mutations produce identical phenotypes affecting the SNa motor nerve. The genes also display dosage-dependent interactions, suggesting that Ptp52F regulates Trn signaling in SNa motor neurons.

Additional Information

© 2009 American Society for Microbiology. Received 18 November 2008; returned for modification 2 January 2009; accepted 19 March 2009. We thank Allen Laughon for anti-Trn antibodies; Stephen Elledge for the cDNA library; Philip James for yeast vectors and strains; Kathy Keegan for the c-Src plasmid; Tony Hunter for a v-Src clone; Amy Cording (Zinn lab) for fixed embryos, discussions, and communication of results before publication; Ed Silverman (Zinn lab) for help with GST fusion protein purifications; and Girish Ratnaparkhi for S2 cells, vectors, protocols, and discussions. A.R. thanks L. S. Shashidhara and the Biology Division of IISER for the use of their facilities. This study was supported by National Institutes of Health grant RO1 NS28182 to K.Z.

Attached Files

Published - Bugga2009p4424Mol_Cell_Biol.pdf

Supplemental Material - Supplementary_Figure_Legend.zip

Supplemental Material - sup_fig_1.zip


Files (1.6 MB)
Name Size Download all
700.1 kB Preview Download
4.9 kB Preview Download
865.9 kB Preview Download

Additional details

August 20, 2023
October 18, 2023