Identifying the molecular mechanisms behind reproductive isolation between closely related yeast species provides avaluable understanding of their evolution. Sequence divergence and chromosomal rearrangements are the main post-zygotic barriers behind reproductive isolation within Saccharomyces „sensu strico’ species, where hybrids are readily formed but sterile upon meiosis. Saccharomyces paradoxus and Saccharomyces cariocanus have an almost identical genome in terms of sequence, and therefore provide good model systems to explore the impact of karyotypic rearrangements on reproductive isolation. According to the biological species concept they are considered two different species despite having low sequence divergence. Since the karyotypic analysis revealed that the genomic differences are restricted to four chromosomal translocations, we hypothesized that such rearrangements may be the cause of low spore viability between them. To test this expectation, we engineered two chromosomal translocations in S. paradoxus YPS138, via Cre-loxP mediated recombination event, to render those parts of genome collinear to S.cariocanus UFRJ50816. Our analysis revealed that hybrids between S. cariocanus and engineered S. paradoxus harbouring two translocations showed a significant increase in spore viability (12.7%) compared to control hybrids harbouring five translocations (3.4%) (P=0.0031and P=0.0125, respectively, Two-sample t-test). Consequently, fitness in meiosis was improved four fold by undoing two translocations. Given this result, the prediction for spore viability in complete collinear crossing would be around 50.8 %, which is still far from the value of ca. 100%, which would be expected for strains with very low sequence divergence and belonging to the same species. This indicates that other factors may contribute to meiotic fitness in these hybrids. Further investigation was carried to determine the genome structures by using the PacBio sequencing approach. Our DNA sequencing data revealed other, previously undetected, rearrangements in S. cariocanus strain: one new reciprocal translocation between chromosomes XIII and XIV and 11 inversions distributed in 6 chromosomes. The variations in meiotic viability observed in the engineered hybrids could be because of these 5 chromosomal translocations. Further experiments were also carried out to evaluate the impact of translocations on mitotic fitness and gene expression; we observed a significant drop in the mitotic fitness of engineered translocant strains under different nutritional and temperature stresses. These changes were also accompanied with alteration in genes expression throughout the genome. Our RNA- seq data revealed that many genes were up- or down- regulated because of the translocation. Several genes with altered expression in translocant strains are correlated with morphology changes when they are up- or down- regulated. Therefore, the cell morphology was evaluated under light microscopy and different abnormal cells were detected compared to the wild type. Irregular cell morphology included elongated and clumped cells. Overall, these data confirmed that chromosomal translocations were the cause of reproductive isolation between S. paradoxus and S. cariocanus and play an important role in altering the phenotype and gene expression.

No description available.

Multiple myeloma is a bone marrow malignancy characterized by the presence of monoclonal plasma cells. In 50-75% of myeloma patients, chromosome translocations at the IgH locus are observed, which result in overexpression of oncogenes from the translocated chromosome due to linkage with the IgH enhancers. IgH enhancer activity is mediated by the B cell-specific transcription factors Bob1 and Oct2. We hypothesized that inhibiting the IgH enhancer, through inhibition of Bob1 and Oct2, is a potential therapeutic strategy for translocation-positive myeloma. The expression and prognostic value of Bob1 and Oct2 in myeloma patient samples were assayed. High Bob1 expression was associated with increased survival, whereas high Oct2 expression was associated with reduced survival. In a t(4;14) myeloma cell line, Bob1 inhibition led to decreased expression of the translocated oncogene, FGFR3; however, this did not lead to decreased proliferation or increased apoptosis. To fully understand the roles of Bob1 and Oct2 in myeloma, further research is required. / Experimental Oncology

The E2A locus is involved in chromosomal translocations associated with acute lymphoblastic leukemia. The most common of these involves a translocation between chromosomes 1 and 19 (t1;19), resulting in expression of the chimeric oncoprotein E2A-PBX1. A direct interaction between transcriptional activation domain 1 (AD1) of E2A and KIX domain of the histone acetyltransferase (HAT) /co-activator CBP is required for E2A-PBX1-mediated leukemia induction in mice. This thesis examines the functional consequences of the direct, physical interaction between E2A and CBP, for both proteins. We demonstrate that the interaction between E2A and CBP/p300, as well as another HAT/co-activator, p/CAF, results in acetylation of E2A. Mutagenesis-based mapping studies identify several lysine residues as substrates for acetylation. Of particular interest, a conserved lysine (K34) located within AD1 is acetylated in vitro and in vivo. Substitution of this residue to arginine impairs transcriptional activation of a luciferase reporter while substitution to glutamine, mimicking the acetylation, restores E2A-mediated transcriptional activation. Recent studies have shown that several transcription factors can modulate the intrinsic HAT activity of CBP/p300. We were surprised to find that E2A proteins enhance acetylation of histones by CBP, in vitro and in vivo, in a KIX domain-independent manner. Acetylation of E2A is also not required for stimulation of CBP/p300 histone acetylation. It appears that E2A interacts with the other CBP domains to mediate this effect, presumably through allosteric effects. In summary, we demonstrate that acetylation of E2A plays a role in mediating the transcriptional activation activity of E2A. Furthermore, acetylation of E2A enhances its interaction with CBP/p300, at least in the presence of additional nuclear factors. We show evidence that p/CAF may mediate this effect. Enhancement of CBP/p300 HAT activity by oncogenic E2A-PBX1 proteins in vivo, suggests that some of its leukemia-promoting effects may be due to E2A-induced gain of function effects on CBP/p300. The enhanced interaction between acetylated E2A and CBP/p300, as well as the E2A-mediated stimulation of histone acetyltransferase activity might play a role in the DNA-binding-independent induction of proliferation. / Thesis (Ph.D, Pathology & Molecular Medicine) -- Queen's University, 2007-09-26 13:37:21.905

Haematological cancers like leukemia and lymphoma are characterized by genetic abnormalities, specifically chromosomal translocations. Analyses of the translocation breakpoint regions in patients have shown that some loci in the genome are more susceptible to breakage than others. However, very little is known about the mechanism of generation of many such chromosomal translocations. In the present study, we have attempted to understand the mechanism of fragility of three regions, which are prone to breaks during translocations in follicular lymphoma (FL) and T-cell leukemia. The t(14;18) translocation in FL is one of the most common chromosomal translocations. Most breaks on chromosome 18 are located at the 3’ UTR of the BCL2 gene and are broadly classified into three clusters, namely major breakpoint region (mbr), minor breakpoint cluster region (mcr) and the intermediate cluster region (icr). The RAG complex has been shown to cleave BCL2 mbr by recognizing an altered DNA structure. In the present study, by using a gel based assay, nature of the non-B DNA structure at BCL2 mbr was identified as parallel intramolecular G-quadruplex. Various studies including circular dichroism (CD), mutagenesis, DMS modification assay and 1H NMR showed the presence of three guanine tetrads in the structure. Further, evidence was also found for the formation of such a G-quadruplex structure within mammalian cells. In an effort to characterize the mechanism of fragility of mcr, a unique pattern of RAG cleavage was observed in a sequence dependent manner. Three independent nicks of equal efficiency were generated by RAGs at the cryptic sequence, “CCACCTCT”, at mcr and at a cytosine upstream of it, unlike a single specific nick at the 5’ of heptamer during V(D)J rearrangement. Interestingly, RAG nicking at mcr occured in the presence of both Mg2+ and Mn2+. Using recombination assay, followed by sequencing of the junctions, we find that mcr can recombine with standard RSS in vivo, albeit at a very low frequency. Mutations to this novel motif abolish recombination at the mcr within the cells. In order to determine the prevalence of t(14;18) translocation in the healthy Indian population, nested PCR approach followed by Southern hybridization was used. Results showed 34% prevalence of t(14;18) translocation in the Indian population. Although, no gender based difference was observed, an age dependent increase was found in adults. Further, presence of the t(14;18) transcripts was also detected. The mechanism underlying the fragility of the t(10;14) translocation involving HOX11 gene in T-cell leukemia is not known. Using primer extension assays on a plasmid DNA containing HOX11 breakpoint region, presence of consistent pause sites corresponding to two G-quadruplex forming regions, flanking the patient breakpoints, were detected. These replication blocks were dependent on K+ ions. Native gel shift assays, mutation analysis, S1 nuclease and CD, further revealed formation of intermolecular G-quadruplexes, unlike the BCL2 mbr. Further, sodium bisulfite modification assay indicated the presence of such structures in the genomic DNA within cells. Hence, we propose that two independent G-quadruplex structures formed in the HOX11 gene could interact with each other, thereby resulting in fragility of the intervening sequences, where majority of the patient breakpoints are mapped. Overall, this study has attempted to understand the role of both sequence and structure of DNA, in generating chromosomal fragility during t(14;18) translocation in FL and t(10;14) translocation in T-cell leukemia. These results may facilitate future studies in unraveling the mechanism leading to genomic instability in other lymphoid cancers.

Thesis (PhD (Botany and Zoology))--Stellenbosch University, 2008. / ENGLISH ABSTRACT: The worldwide movement of biota is of substantial concern for the conservation of biodiversity. The movement of species takes place at three different scales. These are translocations of indigenous species within their natural distribution ranges, the translocation of species outside their natural distribution ranges within geopolitical boundaries (i.e. extralimital introductions) and the translocation of species outside geopolitical boundaries and their natural distribution ranges (i.e. extraregional introductions). Ungulates are extensively translocated at each of these scales and each scale of translocation is expected to have different impacts on biodiversity. Ungulates are translocated for conservation purposes such as reintroducing species to places where they have previously gone extinct and to mitigate inbreeding in small, isolated populations. Ungulates are also extensively translocated for economic and recreational reasons, such as for sport hunting. Translocations for sport hunting include indigenous, extralimital and extraregional species. Concerns for translocations of indigenous species are largely for the loss of genetic diversity through the mixing of genetically distinct populations, and concerns for extralimital and extraregional translocations are for impacts on indigenous biodiversity such as through herbivory, competition, hybridization and disease transmission. This thesis investigates the extent of ungulate introductions globally and at a finer resolution in South Africa. It investigates the pathways, drivers and impacts of ungulate introductions and it also investigates the use of surrogates for genetic distinctiveness for advising the translocation of indigenous ungulates. The study finds that ungulate translocations have been extensive and have lead to the homogenization of ungulate assemblages in countries globally and at a quarter-degree grid-cell resolution in South Africa. Zoos were identified as a potential introduction pathway for extraregional ungulates globally and in South Africa extraregional introductions have made ungulate assemblages more different; whereas large numbers of extralimital introductions have made ungulate assemblages more similar. The homogenization of ungulate species in South Africa has increased with time, due to increased numbers of translocations, particularly of extralimital species. In South Africa translocations have most recently been made to high-income areas with high human population density and high livestock density; whereas in the 1960s ungulates were introduced to areas species poor for indigenous ungulates and marginal for livestock. In South Africa, long distance translocations of indigenous species extralimitally has resulted in extensive range expansions of a magnitude greater than predicted range changes as a result of predicted climate change. When the use of surrogates of genetic distinctiveness for advising translocations was investigated for Africa, the East African rift valley was found to be important in delineating genetic distinctiveness and translocations across this feature should be prevented. Major rivers in Africa also showed potential for delineating genetic distinctiveness in ungulates, but relevant phylogeographic data are needed to confirm this. Sufficient evidence for the impacts of non-indigenous ungulates on biodiversity both in South Africa and globally is lacking despite substantial concern for their impacts. It is suggested that to demonstrate the impacts of non-indigenous ungulates exclosure and enclosure experiments should be used and population declines in indigenous species should be shown. / AFRIKAANSE OPSOMMING: Verskuiwing van plante en diere wêreldwyd lei tot groot kommer in gevolge die bewaring van biodiversiteit. Verskuiwings vind op drie skale plaas, naamlik, die verskuiwing van inheemse spesies binne hulle natuurlike verspreidingsgebied, die verskuiwing van spesies buite hul natuurlike verspreidingsgebied maar binne landsgrense (d.w.s. buite limiet verskuiwings), en die verskuiwing van spesies buite hul natuurlike verspreidingsgebied en buite landsgrense (d.w.s. buite grondgebied verskuiwings). Grootskaalse verskuiwing van hoefdiere vind op al drie skale plaas. Verskuiwing op elk van die skale sal na verwagting biodiversiteit verskillend beïnvloed. Hoefdiere word ook vir bewaringsdoeleindes verskuif, bv. die herbevolking van ‘n gebied waar die spesies uitgesterf het, asook om genetiese probleme wat gepaardgaan met klein bevolkingsgroottes te vermy. Hoefdiere word ook verskuif vir ekonomiese- en ontspannings redes, o.a. vir sportjag, en sluit verskuiwings op al drie skale in. Kommer oor verskuiwings binne ‘n spesies se verspreidingsgebied rus hoofsaaklik op die verlies van genetiese diversiteit a.g.v. die vermenging van vorig genetiese eiensoortige bevolkings, terwyl op die ander twee skale kommer hoofsaaklik gebaseer is op die impakte van kompetisie, interteling, predasie, herbivoor effek, en die verspreiding van parasiete op inheemse biodiversiteit. Hierdie proefskrif ondersoek die omvang van hoefdier verskuiwings globaal asook in Suid Afrika. Die verskuiwingsweë, oorsake en impakte van verskuiwings word hier ondersoek, asook die maatstawwe van genetiese eiensoortigheid om advies oor verskuiwings te verskaf. Die bevindings toon op ekstensiewe verskuiwing van hoefdiere wat eenvormigheid van hoefdier spesiesamestellings wêreldwyd bevorder het, sowel as op ‘n kwartgraad skaal in Suid Afrika. Dieretuine is geïdentifiseer as ‘n potensiële bron van buite grondgebied verskuiwings. In Suid Afrika het buite grondgebied verskuiwings hoefdier spesiesamestellings minder eenvormig gemaak, terwyl buite limiet verskuiwings hoefdier samestellings binne Suid Afrika meer eenvormig gemaak het. Eenvormigheid in hoefdier spesiesamestellings het met tyd in Suid Afrika toegeneem as gevolg van ‘n toename in veral buite limiet verskuiwings. In Suid Afrika is verskuiwings mees onlangs na hoë-inkomste gebiede met hoë menslike bevolkingsen veëdigthede gemaak. In die sestiger jare is hoefdiere egter verskuif na gebiede waar min inheemse hoefdiere voorgekom het en wat marginaal was vir veëboerdery. In Suid Afrika het langafstand buite limiet verskuiwings versoorsaak dat die verspreidingsgebiede van sekere hoefdier spesies groter is as wat die geval sal wees met voorspelde klimaatsveranderinge. Waneer surogaat inligting gebruik word om genetiese verskille tussen bevolkings te identifiseer om verskuiwings te adviseer, word die Oos Afrika Rift vallei geïdentifiseer as ‘n belangrike breuk tussen bevolkings. Verskuiwings oor die vallei moet verkieslik nie plaasvind nie. Groot riviere in Afrika speel ‘n soortgelyke rol, maar verdere genetiese inligting is nodig om dit te bevestig. Alhoewel kommer oor die impak van verskuiwings groot en teoreties verdedigbaar is, bestaan daar te min konkrete bewyse vir die impakte in Suid Afrika. Daar word voorgestel dat manipuleringseksperimente uitgevoer moet word om impakte te demonstreer, en dat aandag veral gegee moet word aan die demonstrasie van impakte op inheemse spesies.

The need to curb increased pollution of environmental resources caused by excessive nitrogen (N) fertilizer application and N fertilizer use inefficiencies in wheat (Triticum aestivum) production systems warrants an inexpensive, sustainable, environmentally sound solution, the root system. Wheat germplasm containing the short arm of rye chromosome 1 (1RS) has recently been found to have larger root system sizes in pot experiments in addition to previously documented higher yields and resistance to leaf, stem and yellow rust. These 1RS lines could therefore be useful in wheat breeding efforts targeting superior root system traits for yield improvements as well as environmental and economic benefits. This dissertation evaluated field root biomass production of Pavon 76 and its wheat-rye (1RS) translocations, effects of root biomass on nitrogen use efficiency, and the temporal variation in their root morphological traits and early growth vigor. The translocation 1RS.1BL had 9 and 23 % higher total root biomass than Pavon 76 at jointing and physiological maturity respectively. Root N uptake peaked at the jointing, where it comprised 22-34% of the total plant N uptake and was lowest at physiological maturity for all genotypes. The inclusion of root N uptake reduced the N utilization efficiency and N harvest index by 6-14 and 7-15% respectively, indicating that the use of only the above ground plant parts over estimates these parameters. In pot experiments, the translocation 1RS .1AL had 12 and 39% higher root biomass than Pavon 76 at anthesis and maturity respectively. 1RS.1BL had 38% higher root mass and 16% longer roots than Pavon76 at physiological maturity. This suggests the existence of differences among the genotypes in below ground partitioning of assimilates at peak nutrient demand (anthesis) for grain filling, and also in rates of root decay and senescence. The lack of differences in root morphological traits among genotypes at early growth stages (6-46 days after sowing) indicates that there are minimal differences in early root growth vigor. The 1RS translocations could therefore expand the wheat breeder's tool box in selections for superior root traits for improved NUE without adverse effects on grain yield.

La déficience intellectuelle (DI) est définie par un QI < 70. La DI, répartie en formes non syndromiques et en formes syndromiques, est observée dans 3 % de la population. Des anomalies chromosomiques sont identifiées dans 15 % des DI syndromiques. Les translocations chromosomiques réciproques (TR) apparemment équilibrées sont observées chez 1 individu sur 1000 et seul 6 % des patients avec une TR de novo apparemment équilibrée ont une DI. Plusieurs mécanismes chromosomiques peuvent expliquer la DI syndromique associée à une TR : (i) un microremaniement déséquilibré identifié par l'utilisation de techniques plus résolutives, (ii) la formation d'un gène de fusion, (iii) un effetde position, (iv) la modification d’une région soumise à une empreinte parentale, (v) une interruption d'un gène au niveau d'un ou des deux points de cassure, (vi) une mutation génique sans rapport avec la TR, (vii) ou encore une cause acquise ou multifactorielle. Nous rapportons l'étude de 12 patients avec DI et porteurs d'une TR de novo apparemment équilibrée. L'analyse systématique par puces à ADN de ces individus a été réalisée avec une résolution de 25 kb. Un déséquilibre infracytogénétique au niveau des points de cassure ou ailleurs dans le génome a été observé chez 3/12 patients. Chez les 9 patients sans anomalies sur puces à ADN, nous avons étudié les points de cassure des remaniements de novo apparemment équilibrés. En dehors de la technique de marche sur le chromosome par FISH, deux autres approches ont été mises en oeuvre : (i) l'Array-Painting qui correspond à l'hybridation sur puces à ADN de chacun des dérivés chromosomiques préalablement séparés par Cytométrie en Flux, (ii) et le séquençage haut débit (WGS - Whole Genome Sequencing). Grâce à l'Array-Painting, nous avons identifié (i) chez 2 patients, des interruptions de gènes pouvant expliquer leur phénotype, à savoir les gènes : KIF1A, AUTS2 et EPHA6 ; (ii) et chez 1 patiente, un point de cassure entraînant une dérégulation de la transcription du gène MEF2C. L'étude par WGS a permis (i) chez 1 patiente, de diagnostiquer un déséquilibre plus complexe que celui observé par puce à ADN ; (ii) chez 2 patients, de mettre en évidence unchromothripsis, qui pourrait avoir un impact dans les pathologies constitutionnelles par interruption de gènes et/ou par effet de position ; (iii) et chez 2 autres patients, de caractériser précisément les points de cassure. Ainsi, grâce aux résultats obtenus par ces différentes techniques, plusieurs mécanismes physiopathologiques responsables de DI sont mis en évidence permettant un conseil génétique adéquat. Cependant, aucun mécanisme chromosomique commun ne peut être identifié hormis le chromothripsis observé chez patients. Finalement, ce travail nous permet principalement de comparer les techniques mises en oeuvre qui se sont avérées complémentaires. En conclusion, nous proposons une démarche diagnostique pour explorer un remaniement chromosomique apparemment équilibré chez des patients à phénotype anormal / Intellectual disability (ID) is defined by an IQ <70. ID, observed in 3% of the population, and displays heterogeneous origins, including acquired etiology (toxicologic, pathologic, traumatic) or genetic disorders with non-syndromic and syndromic forms. Numerical or structural chromosomal abnormalities are observed in 15% of patients with ID. Reciprocal balanced chromosomal translocations (RT) are observed in one individual in 1000. However, only 6% of patients carrying a de novo apparently balanced RT present ID. The relation between these balanced rearrangements and ID could be explained by different mechanisms namely (i) subtle rearrangement, (ii) gene fusion, (iii) position effect, (iv) disturbance of parental imprinting, (v) gene disruption at the breakpoints, (vi) mutation in gene unrelated to the translocation, (vii) or acquired or multifactorial cause. We report a chromosomal study of 12 patients with DI and carrying a de novo apparently balanced reciprocal translocation. A systematic analysis by microarrays was performed in all individuals (using a resolution of 25 kb). For three patients, a microdeletion was observed at the breakpoints or elsewhere in the genome. For the 9 remaining cases, we hypothesize that the phenotype is due to a disruption of gene(s) located at the breakpoint(s). In this context, we studied the breakpoints of the apparently balanced de novo rearrangements in these patients. Outside FISH walking, two approaches have been implemented namely Array-Painting, which combines flow chromosome sorting in an attempt to isolate derivative chromosomes from each other and DNA microarrays as well as Whole Genome Sequencing (WGS). Using Array-Painting, we identified (i) in 2 patients, a gene disruptions: in the KIF1A, AUTS2 and EphA6 genes; (ii) and in 1 patient, a breakpoint resulting in deregulation of transcription of the gene MEF2C. The WGS technology has permitted (i) in 1 patient, to diagnose more complex imbalance than that observed by micro-array; (ii) in 2 patients, to show a chromothripsis, (iii) and 2 other patients, to characterize precisely breakpoints. In conclusion, taking together, these results highlight different physiopathological mechanisms responsible for DI allowing adequate genetic counseling. However, no common chromosomal mechanism can be identified except for chromothripsis observed in 2 patients. In addition, this work allows us especially to compare the used techniques which seem to be complementary. Finally, we propose a pipeline to elucidate the etiology of the abnormal phenotype in patients carrying an apparently balanced rearrangement

Au cours de la réplication, la réparation des cassures double brin (CDB) par recombinaison homologue (RH), basée sur la synthèse d’ADN à partir de la chromatide sœur, permet le maintien de la stabilité du génome. La religature d’extrémités (EJ) éloignées de CDB peut quant à elle générer des réarrangements menaçant son intégrité. Nous avons étudié le mécanisme de réparation par EJ en fonction de la distance séparant deux cassures double brin. En utilisant des substrats intra-chromosomiques permettant la mesure de l’efficacité et de la fidélité du EJ après ligature d’extrémités éloignées ou proximales, nous avons mis en évidence l’implication du complexe de cohésion dans l’inhibition du EJ d’extrémités distales. Le complexe de cohésion joue donc un rôle central dans l’interface réplication/réparation ; la cohésion des chromatides sœurs favorise la réparation par RH et permet l’inhibition spécifique du EJ d’extrémités éloignées, probablement en limitant la mobilité de la chromatine endommagée et la formation d’une synapse propice au rapprochement des extrémités. La religature d’extrémités éloignées est également nécessaire aux mécanismes de diversification des gènes des immunoglobulines tels que la recombinaison V(D)J et la commutation de classe. L’étude de souris Rad21+/- a également démontré une implication du complexe de cohésion dans ces mécanismes essentiels à la diversité de l’information génétique. Le complexe de cohésion étant impliqué dans ces mécanismes et dans l’inhibition des réarrangements complexes tels que les translocations et insertions il est un acteur essentiel de la diversité et de la stabilité génomique. / DNA double-strand breaks (DSBs) repair is essential for genome stability/diversity, but can also generate genome rearrangements. Although non-homologous end-joining (NHEJ) is required for genome stability maintenance, the joining of distant double strand ends (DSE) should inexorably lead to genetic rearrangements. We analyzed the efficiency and accurency of close or distal EJ repair. Our data show that global end-joining is more efficient on close ends (34bp) compared to distal ends (3200bp) and that C-NHEJ is favored on close ends, resulting in more accurate outcome, compared to distal ends where more mutagenic A-EJ events takes place. In addition, the joining of distal ends favors the insertion/capture of DNA sequences. These data show only few kb distances between two DSEs are sufficient to jeopardize DSB repair efficiency and accuracy, leading to complex scars at the re-sealed junctions, and cell response is sufficiently sensitive to differently process such distal ends. We next addressed the question of the mechanisms preventing the joining of distant DSE. We show that depletion of the cohesin complex proteins specifically stimulates the end-joining of I-SceI-induced DSBs distant of 3200bp, while the joining of close DSEs (34bp) remained unaffected. Consistently, exome sequencing and cytogenetic analysis revealed that RAD21 ablation generates large chromosome rearrangements and a strong induction of replication stress-induced chromosome fusions. These data reveal a role for the cohesin complex in the protection against profound genome rearrangements arising through ligation of distant DSEs.

Acute myeloid leukemia (AML) is a highly heterogeneous disorder that results from a block in the differentiation of hematopoietic progenitor cells along with uncontrolled proliferation. In approximately 60% of cases, specific recurrent chromosomal aberrations can be identified by modern cytogenetic techniques, and is an important indicator to classify patients into three prognostic categories: favorable, intermediate, and poor risk. Currently, favorable risk patients are usually treated with chemotherapy while poor risk patients receive allogeneic stem cell transplantation. However, the largest subgroup of AML patients (approximately 40%) has no identifiable cytogenetic abnormalities and is classified as intermediate risk. In this special subgroup of patients, a number of studies have demonstrated the relationship between different translocations involving the mixed lineage leukemia (MLL) gene and patient prognosis. The heterogeneity of MLL-rearranged AML is reflected by the identification of more than 70 different fusion partners of this gene and the panel is continuously increasing. The aim of this study is to develop a sensitive molecular profiling test for relevant risk stratification that can help in the decision of treatment and/or follow-up strategy.