Supplementary Materials

Supplementary Materials for:

The kinase activity of the Ser/Thr kinase BUB1 promotes TGF-β signaling

Shyam Nyati, Katrina Schinske-Sebolt, Sethuramasundaram Pitchiaya, Katerina Chekhovskiy, Areeb Chator, Nauman Chaudhry, Joseph Dosch, Marcian E. Van Dort, Sooryanarayana Varambally, Chandan Kumar-Sinha, Mukesh Kumar Nyati, Dipankar Ray, Nils G. Walter, Hongtao Yu, Brian Dale Ross, Alnawaz Rehemtulla*

*Corresponding author. E-mail: alnawaz{at}med.umich.edu

This PDF file includes:

  • Fig. S1. siRNA screening and pathway analysis.
  • Fig. S2. BUB1 mediates TGF-β–dependent SMAD2/3 phosphorylation.
  • Fig. S3. Depletion of BUB1 leads to reduced SMAD3 recruitment to TGFBRI and TGF-β–dependent accumulation of SMAD2 in the nucleus.
  • Fig. S4. Depletion of BUB1 attenuates TGF-β–dependent SMAD2/3-SMAD4 complex formation in A549 cells.
  • Fig. S5. Depletion of BUB1 inhibits TGF-β–mediated EMT in A549 and NCI-H358 cells.
  • Fig. S6. Depletion of BUB1 inhibits TGF-β–mediated migration and invasion of A549 cells.
  • Fig. S7. BUB1 kinase activity mediates TGF-β–dependent phosphorylation and nuclear activity of R-SMAD.
  • Fig. S8. TGFBRI and BUB1 colocalization by TIRF microscopy at 1 and 24 hours and coimmunoprecipitation of His-TGFBRI and Myc-BUB1.
  • Fig. S9. BUB1 kinase activity mediates TGFBRI-TGFBRII interaction.
  • Fig. S10. TGFBRI is not a direct substrate of BUB1 kinase activity.
  • Fig. S11. Wild-type Myc-BUB1 interacts with FL-SMAD2 in HEK293T cells.
  • Fig. S12. SMAD3 is not a direct substrate of BUB1 kinase activity.
  • Legends for tables S1 and S2
  • Table S3. Pathway impact analysis based on the fold induction of the BTR reporter.

[Download PDF]

Technical Details

Format: Adobe Acrobat PDF

Size: 982 KB

Other Supplementary Material for this manuscript includes the following:

  • Table S1 (Microsoft Excel format). Human kinome siRNA screen in A549-BTR and MDA-231-1833–BTR cells.
  • Table S2 (Microsoft Excel format). Hits obtained in A549-BTR and MDA-231-1833–BTR human kinome screen.

[Download Tables S1 and S2]


Citation: S. Nyati, K. Schinske-Sebolt, S. Pitchiaya, K. Chekhovskiy, A. Chator, N. Chaudhry, J. Dosch, M. E. Van Dort, S. Varambally, C. Kumar-Sinha, M. K. Nyati, D. Ray, N. G. Walter, H. Yu, B. D. Ross, A. Rehemtulla, The kinase activity of the Ser/Thr kinase BUB1 promotes TGF-β signaling. Sci. Signal. 8, ra1 (2015).

© 2014 American Association for the Advancement of Science