A collaborative paper with Professor Sungsoo Na’s group at IUPUI is now published at Scientific Reports.
Subcellular domain-dependent molecular hierarchy of SFK and FAK in mechanotransduction and cytokine signaling
Qiaoqiao Wan, ThucNhi TruongVo, Hannah E. Steele, Altug Ozcelikkale, Bumsoo Han, Yingxiao Wang, Junghwan Oh , Hiroki Yokota & Sungsoo Na
Abstract: Focal adhesion kinase (FAK) and Src family kinases (SFK) are known to play critical roles in mechanotransduction and other crucial cell functions. Recent reports indicate that they reside in different microdomains of the plasma membrane. However, little is known about their subcellular domain-dependent roles and responses to extracellular stimuli. Here, we employed uorescence resonance energy transfer (FRET)-based biosensors in conjunction with collagen-coupled agarose gels to detect subcellular activities of SFK and FAK in three-dimensional (3D) settings. We observed that SFK and FAK in the lipid rafts and nonrafts are differently regulated by fluid flow and pro-inflammatory cytokines. Inhibition of FAK in the lipid rafts blocked SFK response to fluid flow, while inhibition of SFK in the non-rafts blocked FAK activation by the cytokines. Ex-vivo FRET imaging of mouse cartilage explants showed that intermediate level of interstitial fluid flow selectively decreased cytokine-induced SFK/FAK activation. These findings suggest that SFK and FAK exert distinctive molecular hierarchy depending on their subcellular location and extracellular stimuli.