Sassano Maria Livia, Derua Rita, Waelkens Etienne, Agostinis Patrizia, van Vliet Alexander R
Cell Death Research and Therapy Group, Department of Cellular and Molecular Medicine, KU Leuven, Leuven, Belgium.
VIB Center for Cancer Biology Research, Leuven, Belgium.
Contact (Thousand Oaks). 2021 Nov 29;4:25152564211052392. doi: 10.1177/25152564211052392. eCollection 2021 Jan-Dec.
We recently reported that the ER stress kinase PERK regulates ER-mitochondria appositions and ER- plasma membrane (ER-PM) contact sites, independent of its canonical role in the unfolded protein response. PERK regulation of ER-PM contacts was revealed by a proximity biotinylation (BioID) approach and involved a dynamic PERK-Filamin A interaction supporting the formation of ER-PM contacts by actin-cytoskeleton remodeling in response to depletion of ER-Ca stores. In this report, we further interrogated the PERK BioID interactome by validating through co-IP experiments the interaction between PERK and two proteins involved in Ca handling and ER-mitochondria contact sites. These included the vesicle associated membrane (VAMP)-associated proteins (VAPA/B) and the main ER Ca pump sarcoplasmic/endoplasmic reticulum Ca ATPase 2 (SERCA2). These data identify new putative PERK interacting proteins with a crucial role in membrane contact sites and Ca signaling further supporting the uncanonical role of PERK in Ca signaling through membrane contact sites (MCSs).
我们最近报道，内质网应激激酶PERK可调节内质网与线粒体的并置以及内质网与质膜（ER-PM）接触位点，这与其在未折叠蛋白反应中的经典作用无关。通过邻近生物素化（BioID）方法揭示了PERK对ER-PM接触的调节作用，该调节作用涉及动态的PERK-细丝蛋白A相互作用，通过肌动蛋白细胞骨架重塑来支持ER-PM接触的形成，以应对内质网钙库的耗竭。在本报告中，我们通过免疫共沉淀实验验证了PERK与两种参与钙处理和内质网与线粒体接触位点的蛋白质之间的相互作用，进一步探究了PERK BioID相互作用组。这些蛋白质包括囊泡相关膜蛋白（VAMP）相关蛋白（VAPA/B）和主要的内质网钙泵肌浆网/内质网钙ATP酶2（SERCA2）。这些数据鉴定出了新的假定的PERK相互作用蛋白，它们在膜接触位点和钙信号传导中起着关键作用，进一步支持了PERK在通过膜接触位点（MCS）进行钙信号传导中的非经典作用。
