CRTC or TORC Pathway

cAMP-regulated transcriptional co-activators TORC1 (CRTC1) and TORC2 (CRTC2) are known to regulate the cyclic-AMP-responsive element-binding (CREB) protein. Rising intracellular cAMP levels lead to de-phosphorylation of CRTC1 and CRTC2, which leads to the translocation of CRTC to the nucleus. CRTC translocation is known to be regulated by AMPK (5'-AMP activated kinase), PKA and SIK. TORC2 and AMPK are potential therapeutic targets for Type II diabetes. DiscoveRx offers novel direct, functional cellular assays for TORC1 (CRTC1) and TORC2 (CRTC2) translocation. No more assay development required; a simple mix-and-read microtiter plate-based assay  as a ready-to-use biosensor cell line is now available!



Features and Benefits
 

  • Simple, HTS-friendly, whole cell assay
  • Non-imaging method to study nuclear translocation of CRTC1 and CRTC2
  • Chemiluminescent detection: Works on any standard luminometer

Applications
  • Measure compound effects on CRTC1 and CRTC2 translocation 
  • Screen for:  sSmall molecules, siRNA
  • Screen/Profile compounds against upstream targets:  AMPK, S1K, CRTC


Assay List

Cell Lines

Target Description Cat. No.
TORC1 (CRTC1) PathHunter® U2OS TORC1 Nuclear Translocation Cell Line 93-0833C3
TORC2 (CRTC2) PathHunter® U2OS TORC2 Nuclear Translocation Cell Line 93-0813C3
TORC3 (CRTC3) PathHunter® U2OS TORC3 Nuclear Translocation Cell Line 93-1120C3

Assay Ready Kits
Target Description Cat. No.
TORC1 (CRTC1) PathHunter® eXpress TORC1 Nuclear Translocation Assay 93-0833E3CP19L
TORC1 (CRTC1) PathHunter® eXpress TORC1 Nuclear Translocation Assay 93-0833E3CP19M
TORC1 (CRTC1) PathHunter® eXpress TORC1 Nuclear Translocation Assay 93-0833E3CP19S
TORC2 (CRTC2) PathHunter® eXpress TORC2 Nuclear Translocation Assay 93-0813E3CP19L
TORC2 (CRTC2) PathHunter® eXpress TORC2 Nuclear Translocation Assay 93-0813E3CP19M
TORC2 (CRTC2) PathHunter® eXpress TORC2 Nuclear Translocation Assay 93-0813E3CP19S


Assay Principle

The PathHunter® Assay Measures TORC1 or TORC2 translocation and binding to TAZ1 protein. The cells have been engineered to express two complementing fragments of β-Gal within different cellular compartments. Full length human CRTC1 (TORC1) gene or CRTC2 (TORC2) is N-terminally tagged with ProLink (PK). TAZ1(transcriptional adaptor zinc-binding domain) is C-terminally tagged with EA and is localized in the cell nucleus. Phosphorylated TORC does not translocate, but the non-phosphorylated TORC translocates into the nucleus and binds to TAZ1 protein. Activation of the pathway initiates TORC1 translocation to the nucleus that forces complementation of the two β-Gal enzyme fragments (ProLink or ProLabel with EA fragment). This action results in the formation of fully complemented-galactosidase enzyme, the activity of which is measured using chemiluminescent substrate.

 

How it Works
 

PathHunter® U2OS TORC Nuclear Translocation Assay