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XB130/Tks5 scaffold protein interaction regulates Src-mediated cell proliferation and survival.

Moodley S, Hui Bai X, Kapus A, Yang B, Liu M - Mol. Biol. Cell (2015)

Bottom Line: Yeast two-hybrid screening suggests that XB130 interacts with another scaffold protein, Tks5.Structure-function studies showed that the fifth SH3 domain of Tks5 binds to the N-terminus of XB130, which contains polyproline-rich motifs.Furthermore, down-regulation of XB130 and/or Tks5 inhibited serum- and growth factor-induced Src activation and downstream phosphorylation of PI3K and Akt.

View Article: PubMed Central - PubMed

Affiliation: Institute of Medical Science, University of Toronto, Toronto, ON M5S 1A8, Canada Latner Thoracic Surgery Research Laboratories, Toronto General Research Institute, University Health Network, Toronto, ON M5G 1L7, Canada.

No MeSH data available.


Related in: MedlinePlus

XB130 and Tks5 regulate Src activation and signaling. (A) Serum-starved BEAS-2B cells treated with either 10% FBS or 50 ng/ml EGF for up to 60 min show increased total protein tyrosine phosphorylation after 10 min. Nontreated (NT) BEAS-2B cells were not stimulated with FBS or EGF after 18-h serum starvation. (B) Down-regulation of XB130 and/or Tks5 inhibited total protein tyrosine phosphorylation, Src Y416, PI3K p85α Y458, and Akt S473 phosphorylation, compared with control BEAS-2B cells treated with 10% FBS or 50 ng/ ml EGF for 10 min. GAPDH is shown as a loading control.
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Figure 7: XB130 and Tks5 regulate Src activation and signaling. (A) Serum-starved BEAS-2B cells treated with either 10% FBS or 50 ng/ml EGF for up to 60 min show increased total protein tyrosine phosphorylation after 10 min. Nontreated (NT) BEAS-2B cells were not stimulated with FBS or EGF after 18-h serum starvation. (B) Down-regulation of XB130 and/or Tks5 inhibited total protein tyrosine phosphorylation, Src Y416, PI3K p85α Y458, and Akt S473 phosphorylation, compared with control BEAS-2B cells treated with 10% FBS or 50 ng/ ml EGF for 10 min. GAPDH is shown as a loading control.

Mentions: As scaffold proteins and Src-tyrosine kinase substrates, we hypothesized that XB130 and Tks5 may modulate cell proliferation and survival through Src-mediated signaling pathways. BEAS-2B cells were treated with normal DMEM containing either 10% fetal bovine serum FBS) or 50 ng/ml EGF for up to 60 min. Western blot analysis showed that tyrosine phosphorylation of BEAS-2B cellular proteins was rapidly enhanced by 10% FBS or 50 ng/ml EGF treatment after a 10-min interval (Figure 7A). Cells were then transfected with XB130 siRNA or Tks5 siRNA or both XB130 and Tks5 siRNA and treated with either 10% FBS or 50 ng/ml EGF for 10 min. Of importance, total cell lysate tyrosine phosphorylation and phosphorylation of Src Y416 (a sign of Src activation) was clearly enhanced in the control and control siRNA cells treated with 10% FBS or 50 ng/ml EGF (Figure 7B). Down-regulation of XB130, Tks5, or both XB130 and Tks5 inhibited the FBS- or EGF-induced tyrosine phosphorylation of cellular proteins and inhibited the phosphorylation of Src Y416 (Figure 7B).


XB130/Tks5 scaffold protein interaction regulates Src-mediated cell proliferation and survival.

Moodley S, Hui Bai X, Kapus A, Yang B, Liu M - Mol. Biol. Cell (2015)

XB130 and Tks5 regulate Src activation and signaling. (A) Serum-starved BEAS-2B cells treated with either 10% FBS or 50 ng/ml EGF for up to 60 min show increased total protein tyrosine phosphorylation after 10 min. Nontreated (NT) BEAS-2B cells were not stimulated with FBS or EGF after 18-h serum starvation. (B) Down-regulation of XB130 and/or Tks5 inhibited total protein tyrosine phosphorylation, Src Y416, PI3K p85α Y458, and Akt S473 phosphorylation, compared with control BEAS-2B cells treated with 10% FBS or 50 ng/ ml EGF for 10 min. GAPDH is shown as a loading control.
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Figure 7: XB130 and Tks5 regulate Src activation and signaling. (A) Serum-starved BEAS-2B cells treated with either 10% FBS or 50 ng/ml EGF for up to 60 min show increased total protein tyrosine phosphorylation after 10 min. Nontreated (NT) BEAS-2B cells were not stimulated with FBS or EGF after 18-h serum starvation. (B) Down-regulation of XB130 and/or Tks5 inhibited total protein tyrosine phosphorylation, Src Y416, PI3K p85α Y458, and Akt S473 phosphorylation, compared with control BEAS-2B cells treated with 10% FBS or 50 ng/ ml EGF for 10 min. GAPDH is shown as a loading control.
Mentions: As scaffold proteins and Src-tyrosine kinase substrates, we hypothesized that XB130 and Tks5 may modulate cell proliferation and survival through Src-mediated signaling pathways. BEAS-2B cells were treated with normal DMEM containing either 10% fetal bovine serum FBS) or 50 ng/ml EGF for up to 60 min. Western blot analysis showed that tyrosine phosphorylation of BEAS-2B cellular proteins was rapidly enhanced by 10% FBS or 50 ng/ml EGF treatment after a 10-min interval (Figure 7A). Cells were then transfected with XB130 siRNA or Tks5 siRNA or both XB130 and Tks5 siRNA and treated with either 10% FBS or 50 ng/ml EGF for 10 min. Of importance, total cell lysate tyrosine phosphorylation and phosphorylation of Src Y416 (a sign of Src activation) was clearly enhanced in the control and control siRNA cells treated with 10% FBS or 50 ng/ml EGF (Figure 7B). Down-regulation of XB130, Tks5, or both XB130 and Tks5 inhibited the FBS- or EGF-induced tyrosine phosphorylation of cellular proteins and inhibited the phosphorylation of Src Y416 (Figure 7B).

Bottom Line: Yeast two-hybrid screening suggests that XB130 interacts with another scaffold protein, Tks5.Structure-function studies showed that the fifth SH3 domain of Tks5 binds to the N-terminus of XB130, which contains polyproline-rich motifs.Furthermore, down-regulation of XB130 and/or Tks5 inhibited serum- and growth factor-induced Src activation and downstream phosphorylation of PI3K and Akt.

View Article: PubMed Central - PubMed

Affiliation: Institute of Medical Science, University of Toronto, Toronto, ON M5S 1A8, Canada Latner Thoracic Surgery Research Laboratories, Toronto General Research Institute, University Health Network, Toronto, ON M5G 1L7, Canada.

No MeSH data available.


Related in: MedlinePlus