Induction of VEGFA mRNA translation by CoCl2 mediated by HuR.
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.
a Laboratory of Genetics, National Institute on Aging-Intramural Research Program, NIH ; Baltimore , MD USA.|b Laboratory of Cellular and Molecular Neuropharmacology, Department of Drug Sciences, Section of Pharmacology, University of Pavia ; Pavia , Italy.
Cecilia
C
Osera
a Laboratory of Genetics, National Institute on Aging-Intramural Research Program, NIH ; Baltimore , MD USA.
Jennifer L
JL
Martindale
b Laboratory of Cellular and Molecular Neuropharmacology, Department of Drug Sciences, Section of Pharmacology, University of Pavia ; Pavia , Italy.
Marialaura
M
Amadio
a Laboratory of Genetics, National Institute on Aging-Intramural Research Program, NIH ; Baltimore , MD USA.
Jiyoung
J
Kim
a Laboratory of Genetics, National Institute on Aging-Intramural Research Program, NIH ; Baltimore , MD USA.
Xiaoling
X
Yang
a Laboratory of Genetics, National Institute on Aging-Intramural Research Program, NIH ; Baltimore , MD USA.
Christopher A
CA
Moad
a Laboratory of Genetics, National Institute on Aging-Intramural Research Program, NIH ; Baltimore , MD USA.
Fred E
FE
Indig
b Laboratory of Cellular and Molecular Neuropharmacology, Department of Drug Sciences, Section of Pharmacology, University of Pavia ; Pavia , Italy.
Stefano
S
Govoni
a Laboratory of Genetics, National Institute on Aging-Intramural Research Program, NIH ; Baltimore , MD USA.
Kotb
K
Abdelmohsen
a Laboratory of Genetics, National Institute on Aging-Intramural Research Program, NIH ; Baltimore , MD USA.
Myriam
M
Gorospe
b Laboratory of Cellular and Molecular Neuropharmacology, Department of Drug Sciences, Section of Pharmacology, University of Pavia ; Pavia , Italy.
Alessia
A
Pascale
D015687:Cell Hypoxia / Q000235:genetics; D003035:Cobalt / Q000494:pharmacology; D000067780:ELAV-Like Protein 1 / Q000096:biosynthesis / Q000235:genetics*; D005786:Gene Expression Regulation; D006367:HeLa Cells; D006801:Humans; D013058:Mass Spectrometry; D014176:Protein Biosynthesis; D011499:Protein Processing, Post-Translational; D020871:RNA Stability / Q000235:genetics; D012333:RNA, Messenger / Q000235:genetics; D016601:RNA-Binding Proteins / Q000235:genetics; D012261:Ribonucleoproteins / Q000235:genetics* / Q000378:metabolism; D042461:Vascular Endothelial Growth Factor A / Q000096:biosynthesis* / Q000235:genetics
D000067780:ELAV-Like Protein 1; C098770:ELAVL1 protein, human; D012333:RNA, Messenger; D016601:RNA-Binding Proteins; D012261:Ribonucleoproteins; C467484:VEGFA protein, human; D042461:Vascular Endothelial Growth Factor A; D003035:Cobalt; C018021:cobaltous chloride
RNA-binding proteins; biotin pulldown; mass spectrometry; polysome profiling; post-transcriptional gene regulation; ribonucleoprotein complex