Hehnly Lab has a paper at Molecular Biology of the Cell!

by Heidi Hehnly in


Colicino et al. talks about the role of Gravin anchoring PLK1 at mitotic centrosomes. This anchor is important for regulating centrosome organization and function as seen below with super resolution imaging of the centrosome component CEP215 disorganization when Gravin is lost. 

A panel taken from Figure 4 from Colicino et al. of CEP215 disorganization primarily at a single mitotic spindle pole with Gravin loss.  We argue this is through uncontrolled phosphorylation of CEP215 by PLK1 in the absence of Gravin.

A panel taken from Figure 4 from Colicino et al. of CEP215 disorganization primarily at a single mitotic spindle pole with Gravin loss.  We argue this is through uncontrolled phosphorylation of CEP215 by PLK1 in the absence of Gravin.


A collaborative work on PKA and AKAPs

by Heidi Hehnly in


While working in the John Scott lab I had the opportunity to participate in a story with Eileen Kennedy's group entitled PKA-Type I Selective Constrained Peptide Disruptors of AKAP Complexes.   Check it out!

A-Kinase Anchoring Proteins (AKAPs) coordinate complex signaling events by serving as spatiotemporal modulators of cAMP-dependent protein kinase activity in cells. Although AKAPs organize a plethora of diverse pathways, their cellular roles are often elusive due to the dynamic nature of these signaling complexes. AKAPs can interact with the type I or type II PKA holoenzymes by virtue of high-affinity interactions with the R-subunits. As a means to delineate AKAP-mediated PKA signaling in cells, we sought to develop isoform-selective disruptors of AKAP signaling. Here, we report the development of conformationally constrained peptides named RI-STapled Anchoring Disruptors (RI-STADs) that target the docking/dimerization domain of the type 1 regulatory subunit of PKA. These high-affinity peptides are isoform-selective for the RI isoforms, can outcompete binding by the classical AKAP disruptor Ht31, and can selectively displace RIα, but not RIIα, from binding the dual-specific AKAP149 complex. Importantly, these peptides are cell-permeable and disrupt Type I PKA-mediated phosphorylation events in the context of live cells. Hence, RI-STAD peptides are versatile cellular tools to selectively probe anchored type I PKA signaling events.

A-Kinase Anchoring Proteins (AKAPs) coordinate complex signaling events by serving as spatiotemporal modulators of cAMP-dependent protein kinase activity in cells. Although AKAPs organize a plethora of diverse pathways, their cellular roles are often elusive due to the dynamic nature of these signaling complexes. AKAPs can interact with the type I or type II PKA holoenzymes by virtue of high-affinity interactions with the R-subunits. As a means to delineate AKAP-mediated PKA signaling in cells, we sought to develop isoform-selective disruptors of AKAP signaling. Here, we report the development of conformationally constrained peptides named RI-STapled Anchoring Disruptors (RI-STADs) that target the docking/dimerization domain of the type 1 regulatory subunit of PKA. These high-affinity peptides are isoform-selective for the RI isoforms, can outcompete binding by the classical AKAP disruptor Ht31, and can selectively displace RIα, but not RIIα, from binding the dual-specific AKAP149 complex. Importantly, these peptides are cell-permeable and disrupt Type I PKA-mediated phosphorylation events in the context of live cells. Hence, RI-STAD peptides are versatile cellular tools to selectively probe anchored type I PKA signaling events.