MAS1 Receptor Trafficking Involves ERK1/2 Activation Through a β-Arrestin2–Dependent Pathway

Abstract

The MAS1 receptor (R) exerts protective effects in the brain, heart, vessels and kidney. R trafficking plays a critical function in signal termination and propagation and in R resensitization. We examined MAS1R internalization and trafficking upon agonist stimulation and the role of β-arrestin2 in the activation of extracellular signal-regulated kinase 1/2 (ERK1/2) and Akt following MAS1R stimulation. Human embryonic kidney 293T cells were transfected with the coding sequence for MAS1R-YFP. MAS1R internalization was evaluated by measuring the MAS1R present in the plasma membrane after agonist stimulation using a ligand-binding assay. MAS1R trafficking was evaluated by its colocalization with trafficking markers. MAS1R internalization was blocked in the presence of shRNAcaveolin-1 and with dominant negatives for Eps15 (a protein involved in endocytosed Rs by clathrin-coated pits (CCP)) and for dynamin. After stimulation, MAS1R colocalized with Rab11, a slow recycling vesicle marker, and not with Rab4, a fast recycling vesicle marker, or LysoTracker, a lysosome marker. Cells transfected with MAS1R showed an increase in Akt and ERK1/2 activation upon Ang-(1-7) stimulation, which was blocked when the CCP pathway was blocked. Suppression of β-arrestin2 by shRNA reduced the Ang-(1-7)-induced ERK1/2 activation, while Akt activation was not modified. We conclude that upon agonist stimulation, MAS1R is internalized through CCP and caveolae in a dynamin-dependent manner and is then slowly recycled back to the plasma membrane. MAS1R induced Akt and ERK1/2 activation from early endosomes, and the activation of ERK1/2 was mediated by β-arrestin2. Thus, MAS1R activity and density may be tightly controlled by the cell.

COVID-19 and immunomodulation in IBD

“…The classical renin-angiotensin-aldosterone system ACE/angiotensin II/angiotensin type 1/2 receptor (AT1-R) and ACE2/ Ang 1-7/MAS-1 receptor (MAS1-R) systems are shown. [49][50][51][52] The ACE/ AT1-R system has been mainly implicated in pro-inflammatory immune responses and tissue injury. In contrast, the ACE2/MasR system appears to play a key role in many anti-inflammatory pathways controlling tissue protection.…”

“…The ACE2 receptor has been shown to be part of the reninangiotensin-aldosterone system (RAAS) that comprises the classical RAAS ACE/angiotensin II/angiotensin type 1/2 receptor (AT1-R) and ACE2/Ang 1-7/MAS-1 receptor (MAS1-R). [49][50][51][52] While the ACE/AT1-R system has been mainly implicated in proinflammatory immune responses and tissue injury, the ACE2/ MasR system plays a crucial role in many anti-inflammatory pathways controlling tissue protection (figure 2). Functional studies in murine experimental colitis models indicated that modulation of ACE2 expression may affect the severity of colitis activity.…”

Low Expression of Mas1 Receptor in Pancreas Related with Acute Pancreatitis

“…Mas1 receptor exerts protective effects in the brain, heart, vessels, and kidney, and its tissue distribution is ubiquitous [27] . Its up-regulate expression could inhibite autophagy activity in multiple cells and animals.…”

Angiotensin Receptors Heterodimerization and Trafficking: How Much Do They Influence Their Biological Function?

“…Regarding MasR, upon Ang-(1-7) stimulation, MasR is endocyted by CCP and caveolae in a mechanism dependent on dynamin, and then the receptor is directed to the cell surface by slow recycling vesicles (Cerniello et al, 2017). This mechanism of internalization was observed in transfected HEK293T cells and in cultured brainstem neurons from Sprague-Dawley rats, Wistar-Kyoto rats, and spontaneously hypertensive rats (SHRs) (Cerniello et al, 2017;Cerniello et al, 2019).…”

“…Regarding MasR, upon Ang-(1-7) stimulation, MasR is endocyted by CCP and caveolae in a mechanism dependent on dynamin, and then the receptor is directed to the cell surface by slow recycling vesicles (Cerniello et al, 2017). This mechanism of internalization was observed in transfected HEK293T cells and in cultured brainstem neurons from Sprague-Dawley rats, Wistar-Kyoto rats, and spontaneously hypertensive rats (SHRs) (Cerniello et al, 2017;Cerniello et al, 2019). The interesting observation is that MasR undergoes a unique trafficking in brainstem neurons from SHR: The number of MasRs internalized through caveolae was greater compared to that internalized by CCP, and the number of receptors that were returned to the cell membrane was smaller, resulting in a lower amount of resensitized MasRs present at the plasma membrane.…”