Functional interaction of reverse gyrase with single-strand binding protein of the archaeon Sulfolobus

Reverse gyrase is a unique hyperthermophile-specific DNA topoisomerase that induces positive supercoiling. It is a modular enzyme composed of a topoisomerase IA and a helicase domain, which cooperate in the ATP-dependent positive supercoiling reaction. Although its physiological function has not been determined, it can be hypothesized that, like the topoisomerase–helicase complexes found in every organism, reverse gyrase might participate in different DNA transactions mediated by multiprotein complexes. Here, we show that reverse gyrase activity is stimulated by the single-strand binding protein (SSB) from the archaeon Sulfolobus solfataricus. Using a combination of in vitro assays we analysed each step of the complex reverse gyrase reaction. SSB stimulates all the steps of the reaction: binding to DNA, DNA cleavage, strand passage and ligation. By co-immunoprecipitation of cell extracts we show that reverse gyrase and SSB assemble a complex in the presence of DNA, but do not make stable protein–protein interactions. In addition, SSB stimulates reverse gyrase positive supercoiling activity on DNA templates associated with the chromatin protein Sul7d. Furthermore, SSB enhances binding and cleavage of UV-irradiated substrates by reverse gyrase. The results shown here suggest that these functional interactions may have biological relevance and that the interplay of different DNA binding proteins might modulate reverse gyrase activity in DNA metabolic pathways.     

A trade-off between nitrogen uptake and use increases responsiveness to elevated CO2 in infrequently cut mixed C3 grasses

The aim of this study was to evaluate whether the responsiveness of mixed C3 grass species to elevated CO2 is related more to nitrogen uptake or to N-use efficiency. Nitrogen uptake and whole-plant N-use efficiency were investigated with two binary mixtures: Lolium perenne was mixed either with Festuca arundinacea or with Holcus lanatus. The swards were grown on sand with or without CO2 doubling, and subjected to two cutting frequencies. A C20 alcohol was used as a marker to determine species proportion in the total root mass of the mixtures. The mean residence time of N was calculated from that of 15N-labelled fertilizer. Lolium perenne took up significantly more N per unit root mass than its grass competitors, but its N-use efficiency was lower. Elevated CO2 significantly reduced the N uptake of the three grass species. A trade-off between N capture and use was found, as N-use efficiency and N-uptake rate were negatively correlated. A high N-use efficiency, and conversely low N uptake appeared to favour the responsiveness to elevated CO2 of the infrequently cut grasses.     

The protein kinases tightly bound to DNA are present in normal tissues and in regenerating liver, but strongly decreased in hepatomas

We have previously described in rat liver two protein kinases tightly bound to DNA, one is serine-specific, the other arginine-specific. In this work we show that both enzymes are present in various rat tissues and in liver from various species. Both kinase specific activities are strongly decreased in methyl-DBA-induced hepatomas and in HTC cells but not in regenerating liver after hepatectomy. This decrease is then not related to cell proliferation.     

Eveningness is associated with higher risk-taking in dangerous driving situations

Inclination toward eveningness is often associated with risky behavior. But the existing studies are scarce, inconsistent and usually limited to self-reported measures. We sought to investigate in young adults whether morningness-eveningness is associated with risky behavior in dangerous driving situations, with self-reported drunk driving and with alcohol consumption. Results show that, indeed, inclination toward eveningness is associated with these risky behaviors. We also demonstrate a link between morningness-eveningness and sensation seeking. Therefore, young adults with a tendency toward eveningness might be more at risk to face negative consequences of alcohol abuse or to be involved in a road accident.     

Semen quality of 4480 young cancer and systemic disease patients: baseline data and clinical considerations

Background

Except for testicular cancer and Hodgkin’s disease, baseline data on semen quality in case of cancers as well as systemic pathologies of the young adult are scarce or based on low sample size.

Methods

Semen quality in patients having testicular cancer (TGCT, n?=?2315), Hodgkin’s disease (HD, n?=?1175), non-Hodgkin’s lymphoma (NHL, n?=?439), leukemia (L, n?=?360), sarcoma (S, n?=?208), brain tumour (BT, n?=?40), Behcet’s disease (Behcet’s, n?=?68) or multiple sclerosis (MS, n?=?73) was studied and compared to that of 1448 fertile men candidates for sperm donation (CSD) and 208 partners of pregnant women (PPW). All samples were studied following the same methodology in a single laboratory. Post freezing and thawing semen characteristics were also studied.

Results

The percentage of normozoospermic men was only 37 % for L patients and lower than 60 % for TGCT, NHL, S and BT. The level of sperm production was differently decreased according to pathologies, the median total sperm count in TC and L patients being four times lower (p?<?0.01 when compared to CSD and PPW). The lowest percentage of progressively motile spermatozoa was found for L and BT patients (both, p?<?0.01 compared to CSD and PPW). The percentage of morphologically normal spermatozoa was also reduced in cancer patients, especially in BT patients. Progressive motility after thawing in patients was about half that observed among candidates for sperm donation. In almost half of the semen of patients with testicular cancer or leukemia, the total number of motile spermatozoa per straw was less than 0.5?×?10⁶ compared to 4.3?×?10⁶ in CSD.

Conclusions

The present data confirm on large series the deleterious impact of various cancers of the young adult on semen quality, establishing thus baseline data for future studies. Owing to the post-thaw quality of the frozen straws, future fertility projects for the majority of the patients studied (in case there is no post-treatment recovery of spermatogenesis) should necessitate an ICSI to provide the best chance of paternity whatever the fertility check-up in the female partner.

     

Evolution in heterogeneous environments: between soft and hard selection

Since their first formulations about half a century ago, the soft and hard selection models have become classical frameworks to study selection in subdivided populations. These models differ in the timing of density regulation and represent two extreme types of selection: density- and frequency-dependent selection (soft) and density- and frequency-independent selection (hard). Yet only few attempts have been made so far to model intermediate scenarios. Here, we design a model where migration may happen twice during the life cycle: before density regulation with probability d(J) (juvenile migration) and after density regulation with probability d(A) (adult migration). In the first step, we analyze the conditions for the coexistence of two specialists. We find that coexistence is possible under a large range of selection types, even when environmental heterogeneity is low. Then, we investigate the different possible outcomes obtained through gradual evolution. We show that polymorphism is more likely to evolve when the trade-off is weak, environmental heterogeneity is high, migration is low, and in particular when juvenile migration is low relative to adult migration, because the timing of migration affects the magnitude of frequency-dependent selection relative to gene flow. This model may provide a more general theoretical framework to experimentally study evolution in heterogeneous environments.     

Epigenetic inactivation of the miR-124-1 in haematological malignancies

miR-124-1 is a tumour suppressor microRNA (miR). Epigenetic deregulation of miRs is implicated in carcinogenesis. Promoter DNA methylation and histone modification of miR-124-1 was studied in 5 normal marrow controls, 4 lymphoma, 8 multiple myeloma (MM) cell lines, 230 diagnostic primary samples of acute myeloid leukaemia (AML), acute lymphoblastic leukaemia (ALL), chronic myeloid leukaemia (CML), chronic lymphocytic leukaemia (CLL), MM, and non-Hodgkin's lymphoma (NHL), and 53 MM samples at stable disease or relapse. Promoter of miR-124-1 was unmethylated in normal controls but homozygously methylated in 4 of 4 lymphoma and 4 of 8 myeloma cell lines. Treatment of 5-Aza-2'-deoxycytidine led to miR-124-1 demethylation and re-expression of mature miR-124, which also associated with emergence of euchromatic trimethyl H3K4 and consequent downregulation of CDK6 in myeloma cells harboring homozygous miR-124-1 methylation. In primary samples at diagnosis, miR-124-1 methylation was absent in CML but detected in 2% each of MM at diagnosis and relapse/progression, 5% ALL, 15% AML, 14% CLL and 58.1% of NHL (p<0.001). Amongst lymphoid malignancies, miR-124-1 was preferentially methylated in NHL than MM, CLL or ALL. In primary lymphoma samples, miR-124-1 was preferentially hypermethylated in B- or NK/T-cell lymphomas and associated with reduced miR-124 expression. In conclusion, miR-124-1 was hypermethylated in a tumour-specific manner, with a heterochromatic histone configuration. Hypomethylation led to partial restoration of euchromatic histone code and miR re-expression. Infrequent miR-124-1 methylation detected in diagnostic and relapse MM samples showed an unimportant role in MM pathogenesis, despite frequent methylation found in cell lines. Amongst haematological cancers, miR-124-1 was more frequently hypermethylated in NHL, and hence warrants further study.