Skip to main content
. Author manuscript; available in PMC: 2012 Jun 26.
Published in final edited form as: Neuron. 2011 Jun 23;70(6):1085–1099. doi: 10.1016/j.neuron.2011.04.025

Figure 6. Calcineurin-dynamin1 interaction is mediated by a PxIxIT motif found in specific dynamin1 isoforms.

Figure 6

(A) Schematic of PxIxIT box consensus sequence found in the regulatory domain of NFAT (1–4) transcription factors and the PRITIS sequence in the proline-rich domain (PRD) of dynamin1. TAD is the transactivation domain, DNA BD is the DNA binding domain, var-CTD is the variable C-terminal domain for NFAT. PH is the pleckstrin homology domain and GED is the GTPase effector domain for dynamin. (B) Calcineurin-dynamin1 interaction is dependent on the PxIxIT motif. VIVIT peptide (a PxIxIT box mimic), but not a control VEET peptide, blocks association of CaNA with dynamin1. Pulldown with GST alone is shown as control. (C) Calcineurin-dynamin1 interaction via the PxIxIT motif is required for NGF-dependent TrkA internalization. Cell surface biotinylation assay shows that VIVIT, but not VEET treatment decreases NGF-dependent internalization of TrkA receptors. Supernatants were probed for p85. (D) Densitometric quantification of internalized TrkA, **p<0.01, n=4 (E, F) Calcineurin interaction is specific to dynamin1 variants with a PxIxIT box. (E) Schematic of dynamin1 splicing variants. Red box indicates xIxIS portion of the PRITIS box sequence, only present in b tail isoforms. (F) GST pull-down assays with HEK293 lysates show that calcineurin interacts with dynamin1ab via the PxIxIT box, but not dynamin1aa isoforms. HEK293 cells were transfected with dynamin1aa-EGFP, dynamin1ab-EGFP, or dynamin1ab-EGFP with PRITIS sequence mutated to ARATAA.