Noonan and LEOPARD syndromes are developmental disorders with overlapping features, including cardiac abnormalities, short stature and facial dysmorphia. Increased RAS signaling owing to PTPN11, SOS1 and KRAS mutations causes approximately 60% of Noonan syndrome cases, and PTPN11 mutations cause 90% of LEOPARD syndrome cases. Here, we report that 18 of 231 individuals with Noonan syndrome without known mutations (corresponding to 3% of all affected individuals) and two of six individuals with LEOPARD syndrome without PTPN11 mutations have missense mutations in RAF1, which encodes a serine-threonine kinase that activates MEK1 and MEK2. Most mutations altered a motif flanking Ser259, a residue critical for autoinhibition of RAF1 through 14-3-3 binding. Of 19 subjects with a RAF1 mutation in two hotspots, 18 (or 95%) showed hypertrophic cardiomyopathy (HCM), compared with the 18% prevalence of HCM among individuals with Noonan syndrome in general. Ectopically expressed RAF1 mutants from the two HCM hotspots had increased kinase activity and enhanced ERK activation, whereas non-HCM-associated mutants were kinase impaired. Our findings further implicate increased RAS signaling in pathological cardiomyocyte hypertrophy.

N-myristoylation is a common form of co-translational protein fatty acylation resulting from the attachment of myristate to a required N-terminal glycine residue.1,2 We show that aberrantly acquired N-myristoylation of SHOC2, a leucine-rich repeat-containing protein that positively modulates RAS-MAPK signal flow,3–6 underlies a clinically distinctive condition of the neuro-cardio-facial-cutaneous disorders family. Twenty-five subjects with a relatively consistent phenotype previously termed Noonan-like syndrome with loose anagen hair [OMIM 607721]7 shared the 4A>G missense change (Ser2Gly) in SHOC2 that introduces an N-myristoylation site, resulting in aberrant targeting of SHOC2 to the plasma membrane and impaired translocation to the nucleus upon growth factor stimulation. Expression of SHOC2S2G in vitro enhanced MAPK activation in a cell type-specific fashion. Induction of SHOC2S2G in Caenorhabditis elegans engendered protruding vulva, a neomorphic phenotype previously associated with aberrant signaling. These results document the first example of an acquired N-terminal lipid modification of a protein causing human disease.

Deformed wing virus (DWV) of honeybees (Apis mellifera) is closely associated with characteristic wing deformities, abdominal bloating, paralysis, and rapid mortality of emerging adult bees. The virus was purified from diseased insects, and its genome was cloned and sequenced. The genomic RNA of DWV is 10,140 nucleotides in length and contains a single large open reading frame encoding a 328-kDa polyprotein. The coding sequence is flanked by a 1,144-nucleotide 5 nontranslated leader sequence and a 317-nucleotide 3 nontranslated region, followed by a poly(A) tail. The three major structural proteins, VP1 (44 kDa), VP2 (32 kDa), and VP3 (28 kDa), were identified, and their genes were mapped to the N-terminal section of the polyprotein. The C-terminal part of the polyprotein contains sequence motifs typical of well-characterized picornavirus nonstructural proteins: an RNA helicase, a chymotrypsin-like 3C protease, and an RNA-dependent RNA polymerase. The genome organization, capsid morphology, and sequence comparison data indicate that DWV is a member of the recently established genus Iflavirus.Deformed wing virus (DWV) is the one of the main viruses associated with the collapse of honeybee (Apis mellifera L.) colonies due to infestation with the ectoparasitic mite Varroa destructor (4,8,12). The virus was first isolated from a sample of symptomatic honeybees from Japan in the early 1980s and is currently distributed worldwide, wherever varroa mites are found (2, 18). A recent survey of adult bee populations detected DWV in over 90% of French apiaries (66) and in 100% of mite samples. The incidence was slightly reduced when pupal samples were analyzed, especially in the spring (66). DWV has also been detected by serology in the dwarf bee A. florae Fabr. (F. R. Hunter-Fujita, M. S. Mossadegh, and B. V. Ball, Abstr. 36th "Apimondia" Int. Apic. Congr., abstr. 230, 1999) and in the Asian honeybee A. cerana Fabr. (2) and by reverse transcriptase (RT) PCR in bumblebees (29). It is serologically related to Egypt bee virus (8, 9, 13), first isolated in 1977 from infected adults from Egypt (10). Typical symptoms of deformed wing disease are vestigial and crumpled wings, bloated abdomens, paralysis, and a severely shortened adult life span for emerging worker and drone bees (44). For a long time it was believed that these symptoms were due to the feeding activity of the mites (23, 42, 72) until it was shown that, in diseased colonies, deformed bees could emerge from cells not parasitized by varroa mites (49, 55) and that the symptoms can persist in the absence of mites (13). The symptoms are perfectly correlated with the presence of large amounts of DWV, as well as with the reduced virus titers and lower prevalence found in asymptomatic bees from the same colonies (17,19,20,54,67,68). The combination of mites and virus causes immunosuppression in the bees and increased susceptibility to other opportunistic pathogens (76), leading to a progressive reduction in colony performance and a complex disease profile at colony level...