Phosphorylation of seryl-tRNA synthetase by ATM/ATR is essential for hypoxia-induced angiogenesis
Fig 2
SerRS phosphorylation at S101 and S241 inhibits its function in repressing VEGFA expression in cellulo and in vivo.
(A) qRT-PCR analysis of VEGFA expression in HEK293 cells transfected with Flag-tagged SerRSWT or mutants with SerRSAA and SerRSDD substitutions. The exogenous expressions of the SerRS proteins were confirmed by immunoblotting against the Flag tag. VEGFA expression levels are plotted as means ± SEM (n = 3, biological replicates, ****p < 0.0001, Student t test). (B) qRT-PCR analysis of vegfaa transcription in zebrafish injected with SerRS-MO and with co-injection of SerRS-MO and SerRSWT, SerRSAA, or SerRSDD mRNA. Data plotted as means ± SEM (n = 3–4, biological replicates, **p < 0.01, ***p < 0.001, Student t test). The Coomassie blue staining was used to show equal total protein loading. See S1 Data for quantitative data and statistical analysis. See S2 Data for original, uncropped images supporting blots and gel results. qRT-PCR, real-time quantitative reverse transcription PCR; SerRS, seryl-tRNA synthetase; SerRSAA, S101A/S241A; SerRSDD, S101D/S241D; SerRS-MO, antisense morpholino against SerRS; SerRSWT, wild-type SerRS; VEGFA, vascular endothelial growth factor A.