Functional Connection between Rad51 and PML in Homology-Directed Repair

The promyelocytic leukemia protein (PML) is a tumor suppressor critical for formation of nuclear bodies (NBs) performing important functions in transcription, apoptosis, DNA repair and antiviral responses. Earlier studies demonstrated that simian virus 40 (SV40) initiates replication near PML NBs. Here we show that PML knockdown inhibits viral replication in vivo, thus indicating a positive role of PML early in infection. SV40 large T antigen (LT) induces DNA damage and, consequently, nuclear foci of the key homologous recombination repair protein Rad51 that colocalize with PML. PML depletion abrogates LT-induced Rad51 foci. LT may target PML NBs to gain access to DNA repair factors like Rad51 that are required for viral replication. We have used the SV40 model to gain insight to DNA repair events involving PML. Strikingly, even in normal cells devoid of viral oncoproteins, PML is found to be instrumental for foci of Rad51, Mre11 and BRCA1, as well as homology-directed repair after double-strand break (DSB) induction. Following LT expression or external DNA damage, PML associates with Rad51. PML depletion also causes a loss of RPA foci following γ-irradiation, suggesting that PML is required for processing of DSBs. Immunofluorescent detection of incorporated BrdU without prior denaturation indicates a failure to generate ssDNA foci in PML knockdown cells upon γ-irradiation. Consistent with the lack of RPA and BrdU foci, γ-irradiation fails to induce Chk1 activation, when PML is depleted. Taken together, we have discovered a novel functional connection between PML and the homologous recombination-mediated repair machinery, which might contribute to PML tumor suppressor activity

Waste dump erosional landform stability – a critical issue for mountain mining

Mining is the largest producer of solid wastes which, when released to land or into waterways, can cause harmful environmental impacts. This is mostly due to fluvial erosion, which is highly increased in mountain areas, due to abrupt slopes. We have analysed this situation at a mountain watershed (192 ha), where steep mined sites and their waste dumps are the main source of sediment in a Natural Park. This problem was tackled by building gabion check dams downstream from the mined sites. We used the DEM of Differences (DoD) method to quantify erosion and sediment yield from three waste dumps (5 ha). Their topography and substrate properties were analysed to understand the erosion problem. The sediment trapped by the check dams was quantified by Electrical Resistivity Tomography. The rainfall characteristics triggering an episode that filled the check dams with sediment in the winter of 2009-2010, were studied to confirm whether it was a case of extreme precipitation conditions. The waste dumps sediment yield (353 ± 95 Mg ha-1 yr-1) suggests severe landform instability. Analysis of topographic and substrate properties confirmed long, steep slopes combined with highly erodible materials. The check dams proved to be inefficient in controlling sediment loads, as they had only functioned for four years of 31 of existence, having trapped 13000 ± 660 m3 of sediment, whereas we estimated that the waste dumps have yielded approximately three times more sediment for the same period. Rainfall analyses showed that neither intense nor extreme conditions (return period of 25-35 years) triggered the mobilization of 37 ± 2 Mg ha-1 in a month. This study highlights the fact that mining operations in similar mountainous settings, with equivalent waste dump construction and reclamation practices, are currently unfeasible. We conclude that landform stability cannot be achieved at this site without landform changes

Phenotypic and Biomass Yield Variations in Natural Populations of Prairie Cordgrass (Spartina pectinata Link) in the USA

Prairie cordgrass (Spartina pectinata Link) is a productive warm-season, C4 perennial grass native to most of North America having tolerance to wet, cold, and saline growing conditions. Excellent stress tolerance, along with high biomass yields, makes prairie cordgrass a good candidate as a dedicated energy crop on marginal land. However, there is little information available on genetic variation, including yield potential, of native populations in the USA. The objectives of this study were to evaluate biomass yield and to identify the nature and extent of genetic variation in natural populations of prairie cordgrass by comparing endemic strains collected throughout the USA. Forty-two prairie cordgrass populations were collected from prairie-remnant sites in 13 states and evaluated at the University of Illinois in Urbana, IL. The 4-year field study of prairie cordgrass revealed extensive variations in biomass yield and phenotypic traits associated with biomass yield among these populations. Strong correlations were observed between the phenotypic values and origins of the populations. Path coefficient analysis indicated that tiller mass, tiller density, heading date, plant height, and phytomer number positively affected biomass yield directly or indirectly. However, the phenotypic traits including biomass yield showed significant variation among years except for phytomer number and heading date. With the extensive genetic variability and high biomass yield potential demonstrated in this experiment, prairie cordgrass could become a highly productive bioenergy crop by developing a well-planned breeding program

Nuclear organization in genome stability: SUMO connections

Recent findings show that chromatin dynamics and nuclear organization are not only important for gene regulation and DNA replication, but also for the maintenance of genome stability. In yeast, nuclear pores play a role in the maintenance of genome stability by means of the evolutionarily conserved family of SUMO-targeted Ubiquitin ligases (STUbLs). The yeast Slx5/Slx8 STUbL associates with a class of DNA breaks that are shifted to nuclear pores. Functionally Slx5/Slx8 are needed for telomere maintenance by an unusual recombination-mediated pathway. The mammalian STUbL RNF4 associates with Promyelocytic leukaemia (PML) nuclear bodies and regulates PML/PML-fusion protein stability in response to arsenic-induced stress. A subclass of PML bodies support telomere maintenance by the ALT pathway in telomerase-deficient tumors. Perturbation of nuclear organization through either loss of pore subunits in yeast, or PML body perturbation in man, can lead to gene amplifications, deletions, translocations or end-to-end telomere fusion events, thus implicating SUMO and STUbLs in the subnuclear organization of select repair events