Noonan syndrome (NS; MIM 163950) is an autosomal dominant disorder and a member of a family of developmental disorders termed “RASopathies,” which are caused mainly by gain-of-function mutations in genes encoding RAS/MAPK signaling pathway proteins. Whole exome sequencing (WES) and trio-based genomic triangulation of a 15-year-old female with a clinical diagnosis of NS and concomitant cardiac hypertrophy and her unaffected parents identified a de novo variant in
Erin M. Higgins, J. Martijn Bos, Heather Mason-Suares, David J. Tester, Jaeger P. Ackerman, Calum A. MacRae, Katia Sol-Church, Karen W. Gripp, Raul Urrutia, Michael J. Ackerman
ERK activation assay.
Full-length, hemaglutinin-tagged (HA-tagged) WT-MRAS or p.Gly23Val-MRAS was expressed in HEK293T/17 cells. Serum-starved cells were stimulated with EGF (100 ng/ml) and collected in 4× Laemmli sample buffer. Following sonication of the cell lysates, proteins were subjected to electrophoresis and blotted with the indicated antibody. The fraction of ERK that was phosphorylated was calculated for each sample at each time point. (