TMEM16F Regulates Baseline Phosphatidylserine Exposure and Cell Viability in Human Embryonic Kidney Cells

Schenk, Laura K.; Schulze, Ulf; Henke, Sebastian Franz; Weide, Thomas; Pavenstädt, Hermann

doi:10.1159/000445596

Cited by 8 publications

(17 citation statements)

References 33 publications

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“…4e ). An anti-proliferative effect of TMEM16F knockout has also been observed in another study 23 . Moreover, similar to the study by Schenk et al, we observed slightly enhanced phospholipid scrambling after knockdown of TMEM16F and other TMEM16 proteins (Fig.…”

Section: Resultssupporting

confidence: 68%

Contribution of TMEM16F to pyroptotic cell death

et al. 2018

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Pyroptosis is a highly inflammatory form of programmed cell death that is caused by infection with intracellular pathogens and activation of canonical or noncanonical inflammasomes. The purinergic receptor P2X7 is activated by the noncanonical inflammasome and contributes essentially to pyroptotic cell death. The Ca2+ activated phospholipid scramblase and ion channel TMEM16F has been shown earlier to control cellular effects downstream of purinergic P2X7 receptors that ultimately lead to cell death. As pyroptotic cell death is accompanied by an increases in intracellular Ca2+, we asked whether TMEM16F is activated during pyroptosis. The N-terminal cleavage product of gasdermin D (GD-N) is an executioner of pyroptosis by forming large plasma membrane pores. Expression of GD-N enhanced basal Ca2+ levels and induced cell death. We observed that GD-N induced cell death in HEK293 and HAP1 cells, which was depending on expression of endogenous TMEM16F. GD-N activated large whole cell currents that were suppressed by knockdown or inhibition of TMEM16F. The results suggest that whole cell currents induced by the pore forming domain of gasdermin-D, are at least in part due to activation of TMEM16F. Knockdown of other TMEM16 paralogues expressed in HAP1 cells suggest TMEM16F as a crucial element during pyroptosis and excluded a role of other TMEM16 proteins. Thus TMEM16F supports pyroptosis and other forms of inflammatory cell death such as ferroptosis. Its potent inhibition by tannic acid may be part of the anti-inflammatory effects of flavonoids.

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Section: Resultssupporting

confidence: 68%

Contribution of TMEM16F to pyroptotic cell death

et al. 2018

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“…TMEM16B was present at a similarly low level by RT-qPCR. TMEM16F has also been reported in HEK293 [ 51 ]. These channels could be individually or collectively responsible for the current observed.…”

Section: Resultsmentioning

confidence: 99%

CLCA2 is a positive regulator of store-operated calcium entry and TMEM16A

et al. 2018

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The Chloride Channel Accessory (CLCA) protein family was first characterized as regulators of calcium-activated chloride channel (CaCC) currents (ICaCC), but the mechanism has not been fully established. We hypothesized that CLCAs might regulate ICaCC by modulating intracellular calcium levels. In cells stably expressing human CLCA2 or vector, we found by calcium imaging that CLCA2 moderately enhanced intracellular-store release but dramatically increased store-operated entry of calcium upon cytosolic depletion. Moreover, another family member, CLCA1, produced similar effects on intracellular calcium mobilization. Co-immunoprecipitation revealed that CLCA2 interacted with the plasma membrane store-operated calcium channel ORAI-1 and the ER calcium sensor STIM-1. The effect of CLCA2 on ICaCC was tested in HEK293 stably expressing calcium-activated chloride channel TMEM16A. Co-expression of CLCA2 nearly doubled ICaCC in response to a calcium ionophore. These results unveil a new mechanism by which CLCA family members activate ICaCC and suggest a broader role in calcium-dependent processes.

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“…Immunoblotting was carried out as described previously (14,26). Antibodies for ERK, phospho-ERK1/2, GAPDH, phospho-p90 ribosomal S6 kinase (RSK) T359, and RSK1/2/ 3 were from Cell Signaling Technology (Danvers, MA, USA).…”

Section: Western Blotmentioning

confidence: 99%

Angiotensin II regulates phosphorylation of actin‐associated proteins in human podocytes

et al. 2017

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Within the kidney, angiotensin II (AngII) targets different cell types in the vasculature, tubuli, and glomeruli. An important part of the renal filtration barrier is composed of podocytes with their actin-rich foot processes. In this study, we used stable isotope labeling with amino acids in cell culture coupled to mass spectrometry to characterize relative changes in the phosphoproteome of human podocytes in response to short-term treatment with AngII. In 4 replicates, we identified a total of 17,956 peptides that were traceable to 2081 distinct proteins. Bioinformatic analyses revealed that among the increasingly phosphorylated peptides are predominantly peptides that are related to actin filaments, cytoskeleton, lamellipodia, mammalian target of rapamycin, and MAPK signaling. Among others, this screening approach highlighted the increased phosphorylation of actin-bundling protein, L-plastin (LCP1). AngII-dependent phosphorylation of LCP1 in cultured podocytes was mediated by the kinases ERK, p90 ribosomal S6 kinase, PKA, or PKC. LCP1 phosphorylation increased filopodia formation. In addition, treatment with AngII led to LCP1 redistribution to the cell margins, membrane ruffling, and formation of lamellipodia. Our data highlight the importance of AngII-triggered actin cytoskeleton-associated signal transduction in

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TMEM16F Regulates Baseline Phosphatidylserine Exposure and Cell Viability in Human Embryonic Kidney Cells

Cited by 8 publications

References 33 publications

Contribution of TMEM16F to pyroptotic cell death

Contribution of TMEM16F to pyroptotic cell death

CLCA2 is a positive regulator of store-operated calcium entry and TMEM16A

Angiotensin II regulates phosphorylation of actin‐associated proteins in human podocytes

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