Biochemical characterization of novel germline BRAF and MEK mutations in cardio-facio-cutaneous syndrome

Methods Enzymol. 2008:438:277-89. doi: 10.1016/S0076-6879(07)38019-1.

Abstract

Cardio-facio-cutaneous syndrome (CFC) is a sporadic, complex developmental disorder involving characteristic craniofacial features, cardiac defects, ectodermal abnormalities, growth deficiency, hypotonia, and developmental delay. CFC is caused by alteration of activity through the mitogen-activated protein kinase (MAPK) pathway due to heterogeneous de novo germline mutations in B-Raf mutant proteins, MEK1 and MEK2. Approximately 75% of individuals with CFC have mutations in BRAF. In vitro functional studies demonstrate that many of these mutations confer increase activity upon the mutant protein as compared to the wildtype protein. However, as is seen cancer, some of the B-Raf mutant proteins are kinase impaired. Western blot analyses corroborate kinase assays as determined by mutant proteins phosphorylating downstream effectors MEK and ERK. Approximately 25% of individuals with CFC have mutations in either MEK1 or MEK2 that lead to increased MEK kinase activity as judged by increased phosphorylation of its downstream effector ERK. Unlike BRAF, no somatic mutations have ever been identified in MEK genes. The identification of novel germline BRAF and MEK mutations in CFC will help understand the pathophysiology of this syndrome. Furthermore, it will also provide insight to the normal function of B-Raf and MEK, and contribute to the knowledge of the role of the MAPK pathway in cancer. Since the MAPK pathway has been studied intensively in the context of cancer, numerous therapeutics that specifically target this pathway may merit investigation in this population of patients.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Amino Acid Substitution
  • Blotting, Western
  • Craniofacial Abnormalities / genetics*
  • Heart Defects, Congenital / genetics*
  • Humans
  • MAP Kinase Kinase 1 / genetics*
  • MAP Kinase Kinase 2 / genetics*
  • Proto-Oncogene Proteins B-raf / genetics*
  • Skin Abnormalities / genetics*
  • Syndrome
  • raf Kinases / analysis

Substances

  • MAP2K2 protein, human
  • BRAF protein, human
  • Proto-Oncogene Proteins B-raf
  • raf Kinases
  • MAP Kinase Kinase 1
  • MAP Kinase Kinase 2
  • MAP2K1 protein, human