Induction of VEGFA mRNA translation by CoCl2 mediated by HuR

RNA Biology
Cecilia OseraAlessia Pascale

Abstract

Vascular endothelial growth factor (VEGF) A is a master regulator of neovascularization and angiogenesis. VEGFA is potently induced by hypoxia and by pathological conditions including diabetic retinopathy and tumorigenesis. Fine-tuning of VEGFA expression by different stimuli is important for maintaining tissue vascularization and organ homeostasis. Here, we tested the effect of the hypoxia mimetic cobalt chloride (CoCl2) on VEGFA expression in human cervical carcinoma HeLa cells. We found that CoCl2 increased the levels of VEGFA mRNA and VEGFA protein without affecting VEGFA mRNA stability. Biotin pulldown analysis to capture the RNA-binding proteins (RBPs) bound to VEGFA mRNA followed by mass spectrometry analysis revealed that the RBP HuR [human antigen R, a member of the embryonic lethal abnormal vision (ELAV) family of proteins], interacts with VEGFA mRNA. VEGFA mRNA-tagging experiments showed that exposure to CoCl2 increases the interaction of HuR with VEGFA mRNA and promoted the colocalization of HuR and the distal part of the VEGFA 3'-untranslated region (UTR) in the cytoplasm. We propose that under hypoxia-like conditions, HuR enhances VEGFA mRNA translation.

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Citations

Sep 21, 2016·Tumour Biology : the Journal of the International Society for Oncodevelopmental Biology and Medicine·Hao WangYulan Ruan
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Methods Mentioned

BETA
transfection
immunoprecipitation
pulldown
ELISA
RIP
PAR-CLIP
fluorescence microscopy
pull down

Software Mentioned

AxioVision
GraphPad InStat application
Zen
GraphPad
CLIP
PAR

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