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Role of cytoskeletal proteins in cerebral cavernous malformation signaling pathways: a proteomic analysis.

Baxter SS, Dibble CF, Byrd WC, Carlson J, Mack CR, Saldarriaga I, Bencharit S - Mol Biosyst (2014)

Bottom Line: Loss of expression of each CCM gene results in loss of in vitro endothelial tube formation.Principal component analysis and cluster analysis show the effects of individual knockdown.While all CCM mutations result in similar pathology, different CCM mutations have their own distinct pathogenesis in cell signaling.

View Article: PubMed Central - PubMed

Affiliation: David H. Murdock Research Institute, North Carolina Research Campus, Kannapolis, NC 28081, USA.

ABSTRACT
Three genetic mutations were found to cause cerebral cavernous malformation (CCM), a vascular anomaly predisposing affected individuals to hemorrhagic stroke. These CCM proteins function together as a protein complex in the cell. Loss of expression of each CCM gene results in loss of in vitro endothelial tube formation. Label-free differential protein expression analysis using multidimensional liquid chromatography/tandem mass spectrometry (2D-LC-MS/MS) was applied to explore the proteomic profile for loss of each CCM gene expression in mouse endothelial stem cells (MEES) compared to mock shRNA and no shRNA control cell-lines. Differentially expressed proteins were identified (p < 0.05). 120 proteins were differentially expressed among the cell-lines. Principal component analysis and cluster analysis show the effects of individual knockdown. In all knockdown cell-lines, altered expression of cytoskeletal proteins is the most common. While all CCM mutations result in similar pathology, different CCM mutations have their own distinct pathogenesis in cell signaling.

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Summary model for signaling pathway for CCM development.
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fig4: Summary model for signaling pathway for CCM development.

Mentions: The proteomic alteration of cytoskeletal proteins, actin and tubulin isoforms, may be a result of loss of protein complex stability from missing CCM protein interaction. Microtubule stabilization through tubulin–protein interaction is known to regulate the cell polarization and proliferation. For instance, RASSF1A Tumor Suppressor is shown to be involved in the control of microtubule polymerization and in the maintenance of genomic stability.27 This may be similar to CCM complex stabilizing tubulin polymerization through their direct interaction. CCM proteins, e.g. Krit1 and OSM, are also known to interact with actin17,23 and, perhaps stabilize the actin–tubulin complex through their interactions with cofilin and profilin34 (Fig. 4). Cofilin, regulated by LIM-kinase 1,34 controls actin polymerization.35 Cofilin, known as actin-depolymerizing factors, functions with Arp2/3 complex in the opposite direction of profilin to control actin polymerization.36,37 Our proteomic results are inconclusive as to whether cofilin and profilin are differentially expressed among the knockdown, WT, and mock groups. (ESI,† Fig. S1) The relative fold expression of the groups for cofilin and profilin were included as a figure due to previous evidence for the proteins' involvement in the development of the CCM.38


Role of cytoskeletal proteins in cerebral cavernous malformation signaling pathways: a proteomic analysis.

Baxter SS, Dibble CF, Byrd WC, Carlson J, Mack CR, Saldarriaga I, Bencharit S - Mol Biosyst (2014)

Summary model for signaling pathway for CCM development.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4043921&req=5

fig4: Summary model for signaling pathway for CCM development.
Mentions: The proteomic alteration of cytoskeletal proteins, actin and tubulin isoforms, may be a result of loss of protein complex stability from missing CCM protein interaction. Microtubule stabilization through tubulin–protein interaction is known to regulate the cell polarization and proliferation. For instance, RASSF1A Tumor Suppressor is shown to be involved in the control of microtubule polymerization and in the maintenance of genomic stability.27 This may be similar to CCM complex stabilizing tubulin polymerization through their direct interaction. CCM proteins, e.g. Krit1 and OSM, are also known to interact with actin17,23 and, perhaps stabilize the actin–tubulin complex through their interactions with cofilin and profilin34 (Fig. 4). Cofilin, regulated by LIM-kinase 1,34 controls actin polymerization.35 Cofilin, known as actin-depolymerizing factors, functions with Arp2/3 complex in the opposite direction of profilin to control actin polymerization.36,37 Our proteomic results are inconclusive as to whether cofilin and profilin are differentially expressed among the knockdown, WT, and mock groups. (ESI,† Fig. S1) The relative fold expression of the groups for cofilin and profilin were included as a figure due to previous evidence for the proteins' involvement in the development of the CCM.38

Bottom Line: Loss of expression of each CCM gene results in loss of in vitro endothelial tube formation.Principal component analysis and cluster analysis show the effects of individual knockdown.While all CCM mutations result in similar pathology, different CCM mutations have their own distinct pathogenesis in cell signaling.

View Article: PubMed Central - PubMed

Affiliation: David H. Murdock Research Institute, North Carolina Research Campus, Kannapolis, NC 28081, USA.

ABSTRACT
Three genetic mutations were found to cause cerebral cavernous malformation (CCM), a vascular anomaly predisposing affected individuals to hemorrhagic stroke. These CCM proteins function together as a protein complex in the cell. Loss of expression of each CCM gene results in loss of in vitro endothelial tube formation. Label-free differential protein expression analysis using multidimensional liquid chromatography/tandem mass spectrometry (2D-LC-MS/MS) was applied to explore the proteomic profile for loss of each CCM gene expression in mouse endothelial stem cells (MEES) compared to mock shRNA and no shRNA control cell-lines. Differentially expressed proteins were identified (p < 0.05). 120 proteins were differentially expressed among the cell-lines. Principal component analysis and cluster analysis show the effects of individual knockdown. In all knockdown cell-lines, altered expression of cytoskeletal proteins is the most common. While all CCM mutations result in similar pathology, different CCM mutations have their own distinct pathogenesis in cell signaling.

Show MeSH
Related in: MedlinePlus