ADF/cofilin mediates actin cytoskeletal alterations in LLC-PK cells during ATP depletion

Ashworth, Sharon; Southgate, Erica L.; Sandoval, Ruben M.; Meberg, Peter J.; Bamburg, James R.; Molitoris, Bruce A.

doi:10.1152/ajprenal.00210.2002

Cited by 63 publications

(44 citation statements)

References 31 publications

(54 reference statements)

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“…Western blot analysis was used in previous studies to demonstrate ATP depletion-induced dephosphorylation/activation of XAC(wt)-GFP (5). The present studies demonstrated that the levels of phosphorylated XAC(wt)-GFP decreased following 5 min of ATP depletion.…”

Section: Expression Of the Adf/cofilin Isoform Xac(wt)-gfp Leadssupporting

confidence: 56%

“…ATP depletion was accomplished as previously reported (5). Before depletion, confluent MS1 cells were washed three times with HBSS (equilibrated at 37°C with 5% CO 2).…”

Section: Methodsmentioning

confidence: 99%

“…Depleted medium (DMEM without glucose, pyruvate, or amino acids) was added to control plates, and depleted medium containing 0.1 M antimycin A was added to the experimental plates. The cells were returned to the incubator for the ATP depletion periods of 5, 10, 15, 30, and 60 min (5).…”

Section: Methodsmentioning

confidence: 99%

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Cofilin mediates ATP depletion-induced endothelial cell actin alterations

Suurna¹,

Ashworth²,

Hosford³

et al. 2006

American Journal of Physiology-Renal Physiology

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Ischemia and sepsis lead to endothelial cell damage, resulting in compromised microvascular flow in many organs. Much remains to be determined regarding the intracellular structural events that lead to endothelial cell dysfunction. To investigate potential actin cytoskeletal-related mechanisms, ATP depletion was induced in mouse pancreatic microvascular endothelial cells (MS1). Fluorescent imaging and biochemical studies demonstrated a rapid and progressive increase in F-actin along with a decrease in G-actin at 60 min. Confocal microscopic analysis showed ATP depletion resulted in destruction of actin stress fibers and accumulation of F-actin aggregates. We hypothesized these actin alterations were secondary to dephosphorylation/activation of actin-depolymerizing factor (ADF)/cofilin proteins. Cofilin, the predominant isoform expressed in MS1 cells, was rapidly dephosphorylated/activated during ATP depletion. To directly investigate the role of cofilin activation on the actin cytoskeleton during ischemia, MS1 cells were infected with adenoviruses containing the cDNAs for wild-type Xenopus laevis ADF/cofilin green fluorescent protein [XAC(wt)-GFP], GFP, and the constitutively active and inactive isoforms XAC(S3A)-GFP and XAC(S3E)-GFP. The rate and extent of cortical actin destruction and actin aggregate formation were increased in ATP-depleted XAC(wt)-GFP- and XAC(S3A)-GFP-expressing cells, whereas increased actin stress fibers were observed in XAC(S3E)-GFP-expressing cells. To investigate the upstream signaling pathway of ADF/cofilin, LIM kinase 1-GFP (LIMK1-GFP) was expressed in MS1 cells. Cells expressing LIMK1-GFP protein had higher levels of phosphorylated ADF/cofilin, increased stress fibers, and delayed F-actin cytoskeleton destruction during ATP depletion. These results strongly support the importance of cofilin regulation in ischemia-induced endothelial cell actin cytoskeleton alterations leading to cell damage and microvascular dysfunction.

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Section: Expression Of the Adf/cofilin Isoform Xac(wt)-gfp Leadssupporting

confidence: 56%

“…ATP depletion was accomplished as previously reported (5). Before depletion, confluent MS1 cells were washed three times with HBSS (equilibrated at 37°C with 5% CO 2).…”

Section: Methodsmentioning

confidence: 99%

See 1 more Smart Citation

Cofilin mediates ATP depletion-induced endothelial cell actin alterations

Suurna¹,

Ashworth²,

Hosford³

et al. 2006

American Journal of Physiology-Renal Physiology

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“…These proteins are potent regulators of actin filament dynamics in eukaryotic cells and mediate the severing and depolymerization of actin. The actin-depolymerizing factor/cofilin family has been shown to play a crucial role in mediating microvillar breakdown and rapid cytoskeletal alterations in ischemic and ATP-depleted proximal tubule cells (26).…”

mentioning

confidence: 99%

Cofilin Interacts with ClC-5 and Regulates Albumin Uptake in Proximal Tubule Cell Lines

Hryciw

Wang

Devuyst

et al. 2003

Journal of Biological Chemistry

106

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Receptor-mediated endocytosis is a constitutive high capacity pathway for the reabsorption of proteins from the glomerular filtrate by the renal proximal tubule. ClC-5 is a voltage-gated chloride channel found in the proximal tubule where it has been shown to be essential for protein uptake, based on evidence from patients with Dent's disease and studies in ClC-5 knockout mice. To further delineate the role of ClC-5 in albumin uptake, we performed a yeast two-hybrid screen with the Cterminal tail of ClC-5 to identify any interactions of the channel with proteins involved in endocytosis. We found that the C-terminal tail of ClC-5 bound the actin depolymerizing protein, cofilin, a result that was confirmed by GST-fusion pulldown assays. In cultured proximal tubule cells, cofilin was distributed in nuclear, cytoplasmic, and microsomal fractions and co-localized with ClC-5. Phosphorylation of cofilin by overexpressing LIM kinase 1 resulted in a stabilization of the actin cytoskeleton. Phosphorylation of cofilin in two proximal tubule cell models (porcine renal proximal tubule and opossum kidney) was also accompanied by a pronounced inhibition of albumin uptake. This study identifies a novel interaction between the C-terminal tail of ClC-5 and cofilin, an actin-associated protein that is crucial in the regulation of albumin uptake by the proximal tubule.

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“…When ATP levels are not promptly restored, cofilin and ADP actin bind together and form filaments that associate into cylindrical aggregates, called rods. Rods have been observed in the nucleus and cell body of neurons on oxidative stress, 82 glutamate excitotoxicity, 83 and ischemia, 84 and they slowly contribute to neuronal dysfunction and death. Interestingly, the formation of actin rods implies not only an alteration of the synapses but it also contributes to an impairment of gene transcription.…”

Section: Multiple Targets For Gene Repression In the Nucleus Transglumentioning

confidence: 99%

Transglutaminase is a Therapeutic Target for Oxidative Stress, Excitotoxicity and Stroke: A new Epigenetic Kid on the Cns Block

Basso

Ratan

2013

J Cereb Blood Flow Metab

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Transglutaminases (TGs) are multifunctional, calcium-dependent enzymes that have been recently implicated in stroke pathophysiology. Classically, these enzymes are thought to participate in cell injury and death in chronic neurodegenerative conditions via their ability to catalyze covalent, nondegradable crosslinks between proteins or to incorporate polyamines into protein substrates. Accumulating lines of inquiry indicate that specific TG isoforms can shuttle into the nucleus when they sense pathologic changes in calcium or oxidative stress, bind to chromatin and thereby transduce these changes into transcriptional repression of genes involved in metabolic or oxidant adaptation. Here, we review the evidence that supports principally a role for one isoform of this family, TG2, in cell injury and death associated with hemorrhagic or ischemic stroke. We also outline an evolving model in which TG2 is a critical mediator between pathologic signaling and epigenetic modifications that lead to gene repression. Accordingly, the salutary effects of TG inhibitors in stroke may derive from their ability to restore homeostasis by removing inappropriate deactivation of adaptive genetic programs by oxidative stress or extrasynaptic glutamate receptor signaling.

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ADF/cofilin mediates actin cytoskeletal alterations in LLC-PK cells during ATP depletion

Cited by 63 publications

References 31 publications

Cofilin mediates ATP depletion-induced endothelial cell actin alterations

Cofilin mediates ATP depletion-induced endothelial cell actin alterations

Cofilin Interacts with ClC-5 and Regulates Albumin Uptake in Proximal Tubule Cell Lines

Transglutaminase is a Therapeutic Target for Oxidative Stress, Excitotoxicity and Stroke: A new Epigenetic Kid on the Cns Block

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