Proteomics

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Identification of VE-cadherin associated proteins regulated through the lncRNA LASSIE

ABSTRACT: The shear stress-regulated lncRNA LASSIE interacts with cytoskeletal proteins as shown by RNA-antisense purification and subsequent mass spectrometry analysis using an anti-LASSIE and a non-targeting 3’ Desthiobiotin-TEG labeled 2’ O-Me-RNA. As this analysis resulted in high protein binding to the non-targeting control oligonucleotide, the RNA antisense purification was repeated in HUVECs treated with control and anti-LASSIE siRNA. Mass spectrometry analysis confirmed binding of LASSIE to the Intermediate filament protein nestin, as nestin enrichment was decreased in the absence of LASSIE. Additional functional analyses demonstrate a role of LASSIE in shear stress sensing and barrier function in endothelial cells, by stabilizing endothelial cell junctions through connection to the cytoskeleton.

INSTRUMENT(S): Q Exactive

ORGANISM(S): Homo Sapiens (human)

TISSUE(S): Cell Culture

SUBMITTER: Ilka Wittig  

LAB HEAD: Reinier Boon

PROVIDER: PXD018734 | Pride | 2021-09-09

REPOSITORIES: Pride

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Publications

Blood vessels are constantly exposed to shear stress, a biomechanical force generated by blood flow. Normal shear stress sensing and barrier function are crucial for vascular homeostasis and are controlled by adherens junctions (AJs). Here we show that AJs are stabilized by the shear stress-induced long non-coding RNA LASSIE (linc00520). Silencing of LASSIE in endothelial cells impairs cell survival, cell-cell contacts and cell alignment in the direction of flow. LASSIE associates with junction  ...[more]

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