Angiotensin II regulates activation of Bim via Rb/E2F1 during apoptosis: involvement of interaction between AMPKβ1/2 and Cdk4.

Apoptotic cell death is essential for mammalian development and tissue homeostasis. Dysregulation of apoptosis has been identified in pathologies including in pulmonary fibrotic remodeling. We previously reported that a key proapoptotic factor in fibrosis, angiotensin II (Ang II), mediates apoptosis in primary pulmonary artery endothelial cells (PAEC) via the AT(2) receptor and requires activation of AMP-regulated protein kinase (AMPK). We now demonstrate that Ang II induces E2F1 transcription factor binding to and activation of the promoter for the Bcl-2 homology 3 (BH3)-only protein Bim. In PAEC, Ang II treatment induced cyclin-dependent kinase 4 (Cdk4)-mediated hyperphosphorylation of retinoblastoma protein (Rb) and its disassociation from E2F1, a key step in facilitating E2F1-directed transcriptional activity. Indeed, ectopic expression of a dominant negative Cdk4 mutant inhibited Ang II-mediated hyperphosphorylation of Rb and Bim promoter activation. Our data also show that the β-subunit of AMPK was constitutively associated with Cdk4 in PAEC and that Ang II treatment induced AMPKβ phosphorylation and subsequent disassociation of this complex. Both Ang II-induced Rb hyperphosphorylation and Cdk4-AMPK disassociation were blocked by the AMPK inhibitor compound C. Together these findings illuminate a novel proapoptotic signaling pathway in endothelial cells, whereby Ang II triggers E2F1-mediated transcriptional upregulation of Bim via activation of AMPKβ1/2 and Cdk4.