The Cul3-KLHL21 E3 ubiquitin ligase targets aurora B to midzone microtubules in anaphase and is required for cytokinesis.

Cul3 (Cullin3)-based E3 ubiquitin ligases recently emerged as critical regulators of mitosis. In this study, we identify two mammalian BTB (Bric-a-brac-Tramtrack-Broad complex)-Kelch proteins, KLHL21 and KLHL22, that interact with Cul3 and are required for efficient chromosome alignment. Interestingly, KLHL21 but not KLHL22 is necessary for cytokinesis and regulates translocation of the chromosomal passenger complex (CPC) from chromosomes to the spindle midzone in anaphase, similar to the previously described BTB-Kelch proteins KLHL9 and KLHL13. KLHL21 directly binds to aurora B and mediates ubiquitination of aurora B in vitro. In contrast to KLHL9 and KLHL13, KLHL21 localizes to midzone microtubules in anaphase and recruits aurora B and Cul3 to this region. Together, our results suggest that different Cul3 adaptors nonredundantly regulate aurora B during mitosis, possibly by ubiquitinating different pools of aurora B at distinct subcellular localizations.

Comprehensive molecular profiling of lung adenocarcinoma.

Adenocarcinoma of the lung is the leading cause of cancer death worldwide. Here we report molecular profiling of 230 resected lung adenocarcinomas using messenger RNA, microRNA and DNA sequencing integrated with copy number, methylation and proteomic analyses. High rates of somatic mutation were seen (mean 8.9 mutations per megabase). Eighteen genes were statistically significantly mutated, including RIT1 activating mutations and newly described loss-of-function MGA mutations which are mutually exclusive with focal MYC amplification. EGFR mutations were more frequent in female patients, whereas mutations in RBM10 were more common in males. Aberrations in NF1, MET, ERBB2 and RIT1 occurred in 13% of cases and were enriched in samples otherwise lacking an activated oncogene, suggesting a driver role for these events in certain tumours. DNA and mRNA sequence from the same tumour highlighted splicing alterations driven by somatic genomic changes, including exon 14 skipping in MET mRNA in 4% of cases. MAPK and PI(3)K pathway activity, when measured at the protein level, was explained by known mutations in only a fraction of cases, suggesting additional, unexplained mechanisms of pathway activation. These data establish a foundation for classification and further investigations of lung adenocarcinoma molecular pathogenesis.

Non-random autosome segregation: a stepping stone for the evolution of sex chromosome complexes?

A new study in Caenorhabditis elegans shows that homologous autosomes segregate non-randomly with the sex chromosome in the heterogametic sex. Segregation occurs according to size, small autosomes segregating with, and large autosomes segregating away from the X-chromosome. Such sex-biased transmission of autosomes could facilitate the spread of sexually antagonistic alleles whose effects favor the fitness of one sex at the expense of the other. This may provide a first step toward the evolution of new sex determination systems.

Prenatal detection of congenital renal malformations by fetal ultrasonographic examination: an analysis of 709,030 births in 12 European countries.

The study was performed to evaluate the prevalence of prenatal ultrasound diagnoses for renal anomalies in 20 registries of 12 European countries, and to compare the different prenatal scanning policies. Standardized data were acquired from 709,030 livebirths, stillbirths, and induced abortions during the study period of 2.5 years and transmitted for central analysis. At least one renal malformation was diagnosed in 1130 infants and fetuses. Prenatal diagnosis (PD) was given in 81.8% of all cases, 29% of these pregnancies were terminated. The highest detection rate was reported for unilateral multicystic dysplastic kidneys with 97% (102/105). An early diagnosis was documented for exstrophy of bladder at a mean gestational age of 18.5 weeks. Dilatations of the upper urinary tract were seen late in pregnancy at 28.3 weeks. Terminations of pregnancies (TOP) were performed in 67% (58/86) of the detected bilateral renal agenesis/dysgenesis, but only 4% of the unilateral multicystic dysplastic renal malformations (4/102). In about 1/3 of the cases, renal malformations are within the category of associated malformations, which include multiple non-syndromal malformations, chromosomal aberrations, and non-chromosomal syndromes. Renal malformations were detected in 2/3 of the associated category by the first prenatal ultrasound scan. Detection rates vary in the different countries of the European community due to diverse policies, ethical, and religious background. Countries with no routine ultrasound show the lowest rates in detection, and termination of pregnancy. Prenatally detected renal malformations should result in a careful examination for further anomalies. Prenatal ultrasound fulfills the needs of screening examinations and is a good tool in detecting lethal and severe renal malformations.

Two high throughput technologies to detect segmental aneuploidies identify new Williams-Beuren syndrome patients with atypical deletions.

OBJECTIVE: To develop and compare two new technologies for diagnosing a contiguous gene syndrome, the Williams-Beuren syndrome (WBS). METHODS: The first proposed method, named paralogous sequence quantification (PSQ), is based on the use of paralogous sequences located on different chromosomes and quantification of specific mismatches present at these loci using pyrosequencing technology. The second exploits quantitative real time polymerase chain reaction (QPCR) to assess the relative quantity of an analysed locus. RESULTS: A correct and unambiguous diagnosis was obtained for 100% of the analysed samples with either technique (n = 165 and n = 155, respectively). These methods allowed the identification of two patients with atypical deletions in a cohort of 182 WBS patients. Both patients presented with mild facial anomalies, mild mental retardation with impaired visuospatial cognition, supravalvar aortic stenosis, and normal growth indices. These observations are consistent with the involvement of GTF2IRD1 or GTF2I in some of the WBS facial features. CONCLUSIONS: Both PSQ and QPCR are robust, easy to interpret, and simple to set up. They represent a competitive alternative for the diagnosis of segmental aneuploidies in clinical laboratories. They have advantages over fluorescence in situ hybridisation or microsatellites/SNP genotyping for detecting short segmental aneuploidies as the former is costly and labour intensive while the latter depends on the informativeness of the polymorphisms.

Male-pattern baldness susceptibility locus at 20p11.

We conducted a genome-wide association study for androgenic alopecia in 1,125 men and identified a newly associated locus at chromosome 20p11.22, confirmed in three independent cohorts (n = 1,650; OR = 1.60, P = 1.1 x 10(-14) for rs1160312). The one man in seven who harbors risk alleles at both 20p11.22 and AR (encoding the androgen receptor) has a sevenfold-increased odds of androgenic alopecia (OR = 7.12, P = 3.7 x 10(-15)).

Suggestive linkage to chromosome 1q for bone mineral apparent density in Brazilian sister adolescents.

OBJECTIVE: To investigate linkage to chromosome 1q and 11q region for lumbar spine, femoral neck and total body BMD and volumetric BMD in Brazilian sister adolescents aged 10-20-year-old and 57 mothers. METHODS: We evaluated 161 sister pairs (n=329) aged 10-20 years old and 57 of their mothers in this study. Physical traits and lifestyle factors were collected as covariates for lumbar spine (LS), femoral neck (FN) and total body (TB) BMD and bone mineral apparent density (BMAD). We selected nine microsatellite markers in chromosome 1q region (spanning nearly 33cM) and eight in chromosome 11q region (spanning nearly 34cM) to perform linkage analysis. RESULTS: The highest LOD score values obtained from our data were in sister pairs LS BMAD analysis. Their values were: 1.32 (P<0.006), 2.61 (P<0.0002) and 2.44 (P<0.0004) in D1S218, D1S2640 and D1S2623 markers, respectively. No significant LOD score was found with LS and FN BMD/BMAD in chromosome 11q region. Only TB BMD showed significant linkage higher than 1.0 for chromosome 11q region in the markers D11S4191 and D11S937. DISCUSSION/CONCLUSIONS: Our results provided suggestive linkage for LS BMAD at D1S2640 marker in adolescent sister pairs and suggest a possible candidate gene (LHX4) related to adolescent LS BMAD in this region. These results reinforce chromosome 1q21-23 as a candidate region to harbor one or more bone formation/maintenance gene. In the other hand, it did not repeat for chromosome 11q12-13 in our population.

Sex-linked genetic markers in the shrews of the Sorex araneus group : from free gene flow to speciation

SUMMARY: The shrews of the Sorex araneus group are morphologically .very similar, but have undergone a spectacular chromosomal evolution. Altogether, the shrews of this group present a complete array of every possible level of chromosomal and genetic differentiation. In South-Western Europe, four species are recognised: S. antiriorii, S. araneus, S. coronatus and S. granarius, which differ essentially by the amount and the composition of Robertsonian metacentric chromosomés. Additionally, several chromosome races of S. araneus are also present in the same region (i.e. Bretolet, Carlit, Cordon, Jura and Vaud). The objective of this thesis was to examine the genetic relationships between populations, races and /or species of the Sorex araneus group with a special emphasis onsex-specific markers (mtDNA and Y chromosome). We first investigate the evolutionary history of the shrews of the Sorex araneus group distributed in the South-Western Europe. The results of. these analyses confirmed the difficulty to draw a single dichotomic tree within this group. Incongruent mtDNA and Y chromosome phylogenies suggest further that genetic and chromosomal evolution are in this group partially independent processes and that the evolutionary history of the south-western European populations of the S. araneus group can only be understood if we consider secondary contacts between taxa, after their divergence (with genetic exchanges by means of hybridization and / or introgression). Using one male-inherited, one female inherited and eight biparentally inherited markers, we investigate the population genetic structure of the Valais shrew (Sorex antinorii). Overall there results suggest that two already well-differentiated genetic lineages colonized the Swiss Alps after the last glacial period and came into contact in the Rhône Valley. After the Valais shrew (Sorex antinorii) reached the Swiss Alps, it came into contact with the common shrew (Sorex araneus). When two species come into contact and hybridize, endogenous counter-selection of hybrids is usually first expressed as a reduced fertility or viability in hybrids of the heterogametic sex, a mechanism know as Haldane's rule (Haldane 1922). We first evaluated the extent of introgression for Y chromosome, mtDNA and autosomal markers in a hybrid zone between S. antinoriii and S. araneus. The overall level of genetic and karyotypic differentiation between the two species must be strong .enough to allow the detection asymmetric introgression. Secondly, we compared the levels of gene flow between chromosome common to both species and chromosome differently rearranged in each of them. We detected a significantly stronger genetic structure in rearranged chromosomes. Over a 10-year period, we even observed a decrease of genetic structure for common chromosomes. These results strongly support the role of chromosomal rearrangements in the reproductive barrier between S. araneus and S. anfinorii. Overall, this thesis underlines the need to use different inherited (paternally, maternally and / or biparentally) and chromosomally located (on common vs. on rearranged chromosomes) markers to obtain more accurate pictures of genetic relationships between populations or species. RÉSUMÉ: Les musaraignes du groupe Sorex araneus sont morphologiquement très proches, mais ont connu une spectaculaire évolution chromosomique. Prises dans leur ensemble, les musaraignes de ce groupe présentent tous les nivaux possibles de différenciation génétique et chromosomique. Dans le sud-ouest de l'Europe, quatre espèces appartenant à ce groupe sont présentes : S. antinorii, S. araneus, S. coronatus et S. granarius. Celles-ci diffèrent essentiellement par leur caryotype dont la variabilité est principalement due à des fusions Robertsoniennes. De plus, plusieurs races chromosomiques appartenant à S. araneus sont aussi présentes dans la même région (i.e. les races Bretolet, Carlit, Cordon, Jura et Vaud). L'objectif de cette thèse était d'examiner les relations génétiques entre populations, races et/ou espèces du groupe S. araneus, en utilisant particulièrement des marqueurs liés aux sexes (ADN mitochondrial et Chromosome Y). Nous avons dans un premier temps retracé l'histoire évolutive des musaraignes de ce groupe dans le sud-ouest de l'Europe. Les résultats dé ces analyses confirment qu'il est difficile de tracer un simple arbre dichotomique au sein de ce groupe. Les arbres phylogénétiques obtenus sur l'ADN mitochondrial et le chromosome Y sont incongruents et suggèrent de plus que l'évolution génétique et chromosomique sont des processus indépendants. L'histoire évolutive -des populations de ce groupe ne peut. être comprise qu'en considérant des contacts secondaires entre taxa postérieure à leur divergence et induisant des échanges génétiques par hybridation et/ou introgression. Par la suite, nous avons examiné la structure génétique des populations de la musaraigne du Valais, S. antinorii, en utilisant un marqueur transmis par les mâles, un marqueur transmis par les femelles et huit marqueurs transmis par les 2 sexes. Nos résultats suggèrent que deux lignées génétiquement bien différenciées aient colonisé les Alpes Suisses, après les dernières glaciations et entrent en contact dans là Vallée du Rhône. Après avoir franchi les Alpes Suisses, la musaraigne du Valais est entrée en contact avec là musaraigne commune (S. araneus). Lorsque deux espèces entrent en contact et s'hybrident, la sélection contre les hybrides implique habituellement une baisse de fertilité ou de viabilité des hybrides du sexe hétérogamétique (i.e. les mâles XY chez les mammifères). Ce mécanisme est connu sous le nom de règle de Haldane (Haldane 1922) et implique une plus forte structuration génétique de marqueurs males - spécifiques que des marqueurs femelles spécifiques. Nous avons donc évalué le degré d'introgression des marqueurs situés sur le chromosome Y, sur l'ADN mitochondrial et sur des autosomes dans une zone hybride entre S. araneus et S. antinorii. Le niveau de différenciation chromosomique et génétique entre les 2 espèces doit être suffisamment fort pour ne pas permettre la détection d'une introgression asymétrique entre les sexes. Dans un second temps, nous avons comparé les niveaux de flux de gênes mesurés à l'échelle du chromosome, pour des chromosomes communs aux deux espèces et pour des chromosomes différemment arrangées dans chacune des deux espèces. Nous avons détecté une structure génétique significativement plus forte sur les chromosomes réarrangés et comme la zone hybride a été étudiée à dix années d'intervalle, nous observons même une diminution de la structure génétique pour les chromosomes communs au cours du temps.. Ces résultats soutiennent fortement l'hypothèse d'un rôle des réarrangements chromosomiques dans l'établissement d'une barrière reproductive entre S. araneus et S. antinorii. Ainsi cette thèse souligne l'utilité d'utiliser des marqueurs génétiques avec différents modes de transmission. (par les mâles, par les femelles et/ou par les 2 sexes) ou localisés au niveau du chromosome (chromosomes communs vs chromosomes réarrangés) afin d'obtenir une image plus juste ou du moins plus complète des relations génétiques entre populations ou espèces.

Simultaneous Mendelian and clonal genome transmission in a sexually reproducing, all-triploid vertebrate.

Meiosis in triploids faces the seemingly insuperable difficulty of dividing an odd number of chromosome sets by two. Triploid vertebrates usually circumvent this problem through either asexuality or some forms of hybridogenesis, including meiotic hybridogenesis that involve a reproductive community of different ploidy levels and genome composition. Batura toads (Bufo baturae; 3n = 33 chromosomes), however, present an all-triploid sexual reproduction. This hybrid species has two genome copies carrying a nucleolus-organizing region (NOR+) on chromosome 6, and a third copy without it (NOR-). Males only produce haploid NOR+ sperm, while ova are diploid, containing one NOR+ and one NOR- set. Here, we conduct sibship analyses with co-dominant microsatellite markers so as (i) to confirm the purely clonal and maternal transmission of the NOR- set, and (ii) to demonstrate Mendelian segregation and recombination of the NOR+ sets in both sexes. This new reproductive mode in vertebrates ('pre-equalizing hybrid meiosis') offers an ideal opportunity to study the evolution of non-recombining genomes. Elucidating the mechanisms that allow simultaneous transmission of two genomes, one of Mendelian, the other of clonal inheritance, might shed light on the general processes that regulate meiosis in vertebrates.

Heterogeneity of t(4;14) in multiple myeloma. Long-term follow-up of 100 cases treated with tandem transplantation in IFM99 trials.

One hundred de novo multiple myeloma patients with t(4;14) treated with double intensive therapy according to IFM99 protocols were retrospectively analyzed. The median overall survival (OS) and event-free survival (EFS) were 41.4 and 21 months, respectively, as compared to 65 and 37 for patients included in the IFM99 trials without t(4;14) (P<10(-7)). We identified a subgroup of patients presenting at diagnosis with both low beta(2)-microglobulin <4 mg/l and high hemoglobin (Hb) >/=10 g/l (46% of the cases) with a median OS of 54.6 months and a median EFS of 26 months, respectively, which benefits from high-dose therapy (HDT); conversely patients with one or both adverse prognostic factor (high beta(2)-microglobulin and/or low Hb) had a poor outcome. The achievement of either complete response or very good partial response after HDT was also a powerful independent prognostic factor for both OS and EFS.

Sex-chromosome differentiation parallels postglacial range expansion in European tree frogs (Hyla arborea).

Occasional XY recombination is a proposed explanation for the sex-chromosome homomorphy in European tree frogs. Numerous laboratory crosses, however, failed to detect any event of male recombination, and a detailed survey of NW-European Hyla arborea populations identified male-specific alleles at sex-linked loci, pointing to the absence of XY recombination in their recent history. Here, we address this paradox in a phylogeographic framework by genotyping sex-linked microsatellite markers in populations and sibships from the entire species range. Contrasting with postglacial populations of NW Europe, which display complete absence of XY recombination and strong sex-chromosome differentiation, refugial populations of the southern Balkans and Adriatic coast show limited XY recombination and large overlaps in allele frequencies. Geographically and historically intermediate populations of the Pannonian Basin show intermediate patterns of XY differentiation. Even in populations where X and Y occasionally recombine, the genetic diversity of Y haplotypes is reduced below the levels expected from the fourfold drop in copy numbers. This study is the first in which X and Y haplotypes could be phased over the distribution range in a species with homomorphic sex chromosomes; it shows that XY-recombination patterns may differ strikingly between conspecific populations, and that recombination arrest may evolve rapidly (<5000 generations).

LDL-cholesterol concentrations: a genome-wide association study.

BACKGROUND: LDL cholesterol has a causal role in the development of cardiovascular disease. Improved understanding of the biological mechanisms that underlie the metabolism and regulation of LDL cholesterol might help to identify novel therapeutic targets. We therefore did a genome-wide association study of LDL-cholesterol concentrations. METHODS: We used genome-wide association data from up to 11,685 participants with measures of circulating LDL-cholesterol concentrations across five studies, including data for 293 461 autosomal single nucleotide polymorphisms (SNPs) with a minor allele frequency of 5% or more that passed our quality control criteria. We also used data from a second genome-wide array in up to 4337 participants from three of these five studies, with data for 290,140 SNPs. We did replication studies in two independent populations consisting of up to 4979 participants. Statistical approaches, including meta-analysis and linkage disequilibrium plots, were used to refine association signals; we analysed pooled data from all seven populations to determine the effect of each SNP on variations in circulating LDL-cholesterol concentrations. FINDINGS: In our initial scan, we found two SNPs (rs599839 [p=1.7x10(-15)] and rs4970834 [p=3.0x10(-11)]) that showed genome-wide statistical association with LDL cholesterol at chromosomal locus 1p13.3. The second genome screen found a third statistically associated SNP at the same locus (rs646776 [p=4.3x10(-9)]). Meta-analysis of data from all studies showed an association of SNPs rs599839 (combined p=1.2x10(-33)) and rs646776 (p=4.8x10(-20)) with LDL-cholesterol concentrations. SNPs rs599839 and rs646776 both explained around 1% of the variation in circulating LDL-cholesterol concentrations and were associated with about 15% of an SD change in LDL cholesterol per allele, assuming an SD of 1 mmol/L. INTERPRETATION: We found evidence for a novel locus for LDL cholesterol on chromosome 1p13.3. These results potentially provide insight into the biological mechanisms that underlie the regulation of LDL cholesterol and might help in the discovery of novel therapeutic targets for cardiovascular disease.

Detection of CBP rearrangements in acute myelogenous leukemia with t(8;16).

The CREB-binding protein (CBP) is a large nuclear protein that regulates many signal transduction pathways and is involved in chromatin-mediated transcription. The translocation t(8;16)(p11;p13.3) consistently disrupts two genes: the CBP gene on chromosome band 16p13.3 and the MOZ gene on chromosome band 8p11. Although a fusion of these two genes as a result of the translocation is expected, attempts at detecting the fusion transcript by reverse transcriptase polymerase chain reaction (RT-PCR) have proven difficult; to date, only one in-frame CBP/MOZ fusion transcript has been reported. We therefore sought other reliable means of detecting CBP rearrangements. We applied fluorescence in situ hybridization (FISH) and Southern blot analyses to a series of AML patients with a t(8;16) and detected DNA rearrangements of both the CBP and the MOZ loci in all cases tested. All six cases examined for CBP rearrangements have breakpoints within a 13 kb breakpoint cluster region at the 5' end of the CBP gene. Additionally, we used a MOZ cDNA probe to construct a surrounding cosmid contig and detect DNA rearrangements in three t(8;16) cases, all of which display rearrangements within a 6 kb genomic fragment of the MOZ gene. We have thus developed a series of cosmid probes that consistently detect the disruption of the CBP gene in t(8;16) patients. These clones could potentially be used to screen other cancer-associated or congenital translocations involving chromosome band 16p13.3 as well.

Partial trisomy 20q in a newborn with dextrocardia.

A female newborn is reported with dextrocardia and a partial trisomy 20q, derived from a t(2;20) paternal translocation. The most discriminating findings of the condition include brachycephaly, bulging forehead, deep set eyes, short nose, large ears, dimpled chin, short neck and a heart defect. Previously reported patients with this rare chromosomal anomaly are reviewed.