Exome sequencing of extreme phenotypes identifies DCTN4 as a modifier of chronic Pseudomonas aeruginosa infection in cystic fibrosis

Exome sequencing has become a powerful and effective strategy for the discovery of genes underlying Mendelian disorders. However, use of exome sequencing to identify variants associated with complex traits has been more challenging, partly because the sample sizes needed for adequate power may be very large. One strategy to increase efficiency is to sequence individuals who are at both ends of a phenotype distribution (those with extreme phenotypes). Because the frequency of alleles that contribute to the trait are enriched in one or both phenotype extremes, a modest sample size can potentially be used to identify novel candidate genes and/or alleles. As part of the National Heart, Lung, and Blood Institute (NHLBI) Exome Sequencing Project (ESP), we used an extreme phenotype study design to discover that variants in DCTN4, encoding a dynactin protein, are associated with time to first P. aeruginosa airway infection, chronic P. aeruginosa infection and mucoid P. aeruginosa in individuals with cystic fibrosis.     

Mapping autism risk loci using genetic linkage and chromosomal rearrangements

Autism spectrum disorders (ASDs) are common, heritable neurodevelopmental conditions. The genetic architecture of ASDs is complex, requiring large samples to overcome heterogeneity. Here we broaden coverage and sample size relative to other studies of ASDs by using Affymetrix 10K SNP arrays and 1,181 [corrected] families with at least two affected individuals, performing the largest linkage scan to date while also analyzing copy number variation in these families. Linkage and copy number variation analyses implicate chromosome 11p12-p13 and neurexins, respectively, among other candidate loci. Neurexins team with previously implicated neuroligins for glutamatergic synaptogenesis, highlighting glutamate-related genes as promising candidates for contributing to ASDs.     

Oncogenically active MYD88 mutations in human lymphoma

The activated B-cell-like (ABC) subtype of diffuse large B-cell lymphoma (DLBCL) remains the least curable form of this malignancy despite recent advances in therapy. Constitutive nuclear factor (NF)-κB and JAK kinase signalling promotes malignant cell survival in these lymphomas, but the genetic basis for this signalling is incompletely understood. Here we describe the dependence of ABC DLBCLs on MYD88, an adaptor protein that mediates toll and interleukin (IL)-1 receptor signalling, and the discovery of highly recurrent oncogenic mutations affecting MYD88 in ABC DLBCL tumours. RNA interference screening revealed that MYD88 and the associated kinases IRAK1 and IRAK4 are essential for ABC DLBCL survival. High-throughput RNA resequencing uncovered MYD88 mutations in ABC DLBCL lines. Notably, 29% of ABC DLBCL tumours harboured the same amino acid substitution, L265P, in the MYD88 Toll/IL-1 receptor (TIR) domain at an evolutionarily invariant residue in its hydrophobic core. This mutation was rare or absent in other DLBCL subtypes and Burkitt's lymphoma, but was observed in 9% of mucosa-associated lymphoid tissue lymphomas. At a lower frequency, additional mutations were observed in the MYD88 TIR domain, occurring in both the ABC and germinal centre B-cell-like (GCB) DLBCL subtypes. Survival of ABC DLBCL cells bearing the L265P mutation was sustained by the mutant but not the wild-type MYD88 isoform, demonstrating that L265P is a gain-of-function driver mutation. The L265P mutant promoted cell survival by spontaneously assembling a protein complex containing IRAK1 and IRAK4, leading to IRAK4 kinase activity, IRAK1 phosphorylation, NF-κB signalling, JAK kinase activation of STAT3, and secretion of IL-6, IL-10 and interferon-β. Hence, the MYD88 signalling pathway is integral to the pathogenesis of ABC DLBCL, supporting the development of inhibitors of IRAK4 kinase and other components of this pathway for the treatment of tumours bearing oncogenic MYD88 mutations.     

Distribution, foraging behaviour, and capture results of the spotted bat ( Euderma maculatum ) in central Oregon

The spotted bat ( Euderma maculatum ) has been virtually unknown in Oregon despite the existence of potential habitat in many areas of the state. In 2002 and 2003 we searched for spotted bats along the John Day, Deschutes, and Crooked Rivers and at a remote dry canyon southeast of the city of Bend in central Oregon. The species was documented through the use of mist-nets, a bat detector, and recognition of audible spotted bat calls. Spotted bats were found at 11 locations in 6 Oregon counties. Nightly activity patterns of spotted bats were unpredictable. Spotted bats were found in 78% of search areas but on only 48% of survey nights. We observed spotted bats foraging above fields and low upland slopes adjacent to rivers and creeks and along the rims of cliffs. Estimated flying heights of spotted bats ranged from 3 m to 50 m aboveground. The species was difficult to capture and was captured only after considerable experimentation with methods and materials. Three spotted bats were captured toward the end of the project in 2003 and accounted for only 0.5% of all bats captured during the study. Although we attached radio transmitters to 2 spotted bats, we found no roost locations. We believe additional spotted bat surveys in Oregon are warranted, especially in higher-elevation habitats, but recommend that to increase their effectiveness, surveys accommodate the unique foraging behavior of the species.     

The tetracycline resistome

Resistance to tetracycline emerged soon after its discovery six decades ago. Extensive clinical and non-clinical uses of this class of antibiotic over the years have combined to select for a large number of resistant determinants, collectively termed the tetracycline resistome. In order to impart resistance, microbes use different molecular mechanisms including target protection, active efflux, and enzymatic degradation. A deeper understanding of the structure, mechanism, and regulation of the genes and proteins associated with tetracycline resistance will contribute to the development of tetracycline derivatives that overcome resistance. Newer generations of tetracyclines derived from engineering of biosynthetic genetic programs, semi-synthesis, and in particular recent developments in their chemical synthesis, together with a growing understanding of resistance, will serve to retain this class of antibiotic to combat pathogens.     

Allozymes and the hybrid origin of the parthenogenetic lizardCnemidophorus exsanguis

Summary Allozyme electrophoresis was used to determine the hybrid parentage of the triploid parthenogenetic speciesCnemidophorus exsanguis. 19 of 33 loci were polymorphic inC. exsanguis and/or the 7 potential bisexual progenitors. It was determined from allele distributions thatC. exsanguis containes genomes form 3 different species, probablyC. costatus, C. inornatus andC. septemvittatus.Acknowledgments. We thank R.L. Bezy, W.M. Brown, A.E. Greer, C.S. Lieb, J.L. Patton and K.L. Wright for assistance in the field; the Departments of Game and Fish of Arizona and New Mexico and the Direccion General de la Fauna Silvestre, Mexico for providing collecting permits; and R.L. Bezy, H.W. Greene and D.B. Wake for discussions and criticisms. This research was supported in part by NSF Grants DEB 76-20599 and DEB 81-05615 and by a Research Fellowship in Herpetology from the Natural History Museum of Los Angeles County to DAG.