Balanced Production of Ribosome Components Is Required for Proper G1/S Transition in Saccharomyces cerevisiae

Cell cycle regulation is a very accurate process that ensures cell viability and the genomic integrity of daughter cells. A fundamental part of this regulation consists in the arrest of the cycle at particular points to ensure the completion of a previous event, to repair cellular damage, or to avoid progression in potentially risky situations. In this work, we demonstrate that a reduction in nucleotide levels or the depletion of RNA polymerase I or III subunits generates a cell cycle delay at the G₁/S transition in Saccharomyces cerevisiae. This delay is concomitant with an imbalance between ribosomal RNAs and proteins which, among others, provokes an accumulation of free ribosomal protein L5. Consistently with a direct impact of free L5 on the G₁/S transition, rrs1 mutants, which weaken the assembly of L5 and L11 on pre-60S ribosomal particles, enhance both the G₁/S delay and the accumulation of free ribosomal protein L5. We propose the existence of a surveillance mechanism that couples the balanced production of yeast ribosomal components and cell cycle progression through the accumulation of free ribosomal proteins. This regulatory pathway resembles the p53-dependent nucleolar-stress checkpoint response described in human cells, which indicates that this is a general control strategy extended throughout eukaryotes.     

Establishment and survival of the South African legume Lessertia spp. and rhizobia in Western Australian agricultural systems

Background and aims

The South African herbaceous legume species Lessertia capitata, L. diffusa, L. excisa L. incana and L. herbacea were introduced to Australia to assess plant establishment and survival, as well as the saprophytic ability of their root nodule bacteria (RNB).

Methods

Five Lessertia spp., were inoculated with selected RNB strains and were sown in five different agroclimatic areas of the Western Australian wheat-belt during 2007 and 2008. Plant population and summer survival were evaluated in situ. Soil samples and nodules from host plants were also taken from each site. The re-isolated rhizobia were RPO1-PCR fingerprinted and their partial dnaK and nodA genes were sequenced to confirm their identity.

Results

Plants achieved only poor establishment followed by weak summer survival. More than 83 % of the rhizobia re-isolated from Lessertia did not correlate with the original inoculants’ fingerprints, and were identified as Rhizobium leguminosarum. The nodA sequences of the naturalised strains were also clustered with R. leguminosarum sequences, thus eliminating the likelihood of lateral gene transference from Mesorhizobium and suggesting a competition problem with indigenous rhizobia.

Conclusion

The stressful soil conditions and high numbers of resident R. leguminosarum strains in Western Australian soils, and their ability to rapidly nodulate Lessertia spp. but not fix nitrogen are likely to preclude the adoption of Lessertia as an agricultural legume in this region.     

Differences in sexual behavior of two distant populations of the funnel-web wolf spider Aglaoctenus lagotis

Generally, sexual repertoire within a species is conserved, but frequencies of occurrence of sexual behavioral acts often vary, and wide geographic distributions may favor these variations. Aglaoctenus lagotis is distributed along South America and belongs to Sosippinae, a subfamily of wolf spiders that builds funnel webs. Previous reports, based on different populations, suggested variations in sexual behavior and life cycle. Our objective was to describe and compare the sexual behavior of two populations of A. lagotis: ‘Southern Uruguay’ (SU) and ‘Central Argentina’ (CA). We carried out intrapopulation trials, in the laboratory, with 15 female–male pair matings. The most representative courtship acts in SU were web-stretching, striding-forward and forelegs-elevating, whereas in CA they were alternate-waving, web-stretching and leg-tapping. Juddering, forelegs-elevating and striding-forward were exclusive for SU, while alternate-waving and simultaneous-waving were exclusive for CA. We also found differences in copulatory characteristics such as frequencies of insertions and ejaculations. We documented body-shaking during copulation for the first time. Some sexual acts were exclusive of each population, while the shared ones differed in pattern and frequencies of occurrence. In addition to variations in sexual periods, these differences may favor divergence processes or an exceptional high level of flexibility in the sexual behavior of A. lagotis.     

Relatedness does not predict vigilance in a population of the social rodent Octodon degus

The possibility that social foragers adjust and coordinate their scanning activity when in the presence of close relatives to attain inclusive fitness benefits remains controversial and scarcely examined. To this aim, we first tested the null hypothesis of no association between foraging individuals of the diurnal rodent, Octodon degus and their pairwise relatedness (six microsatellite loci), under natural conditions. Secondly, we examined the influence of relatedness on scan effort (percent overlapping) and temporal distribution of scanning using linear regression. Finally, we evaluated whether temporal distributions of scanning were significantly lower (coordination) or higher (synchrony) than random expectations using bootstrapping. We found that pairwise relatedness between focal degus and their foraging partner did not influence the scan effort or the temporal distribution of scanning. These original, field-based findings imply that vigilance behavior in socially foraging degus is unlikely to be kin-selected and adds to results from previous lab studies in that kinship remains a poor predictor of social behavior in these animals. Overall, our study adds to others revealing that kin selection may not have had an impact on aspects of social behavior such as vigilance during social foraging.     

Foot-and-mouth disease virus mutant with decreased sensitivity to ribavirin: implications for error catastrophe

The nucleoside analogue ribavirin (R) is mutagenic for foot-and-mouth disease virus (FMDV). Passage of FMDV in the presence of increasing concentrations of R resulted in the selection of FMDV with the amino acid substitution M296I in the viral polymerase (3D). Measurements of progeny production and viral fitness with chimeric viruses in the presence and absence of R documented that the 3D substitution M296I conferred on FMDV a selective replicative advantage in the presence of R but not in the absence of R. In polymerization assays, a purified mutant polymerase with I296 showed a decreased capacity to use ribavirin triphosphate as a substrate in the place of GTP and ATP, compared with the wild-type enzyme. The results suggest that M296I has been selected because it attenuates the mutagenic activity of R with FMDV. Replacement M296I is located within a highly conserved stretch in picornaviral polymerases which includes residues that interact with the template-primer complex and probably also with the incoming nucleotide, according to the three-dimensional structure of FMDV 3D. Given that a 3D substitution, distant from M296I, was associated with resistance to R in poliovirus, the results indicate that picornaviral polymerases include different domains that can alter the interaction of the enzyme with mutagenic nucleoside analogues. Implications for lethal mutagenesis are discussed.     

Surrogate Markers of Visceral Adiposity in Young Adults: Waist Circumference and Body Mass Index Are More Accurate than Waist Hip Ratio,Model of Adipose Distribution and Visceral Adiposity Index

Surrogate indexes of visceral adiposity, a major risk factor for metabolic and cardiovascular disorders, are routinely used in clinical practice because objective measurements of visceral adiposity are expensive, may involve exposure to radiation, and their availability is limited. We compared several surrogate indexes of visceral adiposity with ultrasound assessment of subcutaneous and visceral adipose tissue depots in 99 young Caucasian adults, including 20 women without androgen excess, 53 women with polycystic ovary syndrome, and 26 men. Obesity was present in 7, 21, and 7 subjects, respectively. We obtained body mass index (BMI), waist circumference (WC), waist-hip ratio (WHR), model of adipose distribution (MOAD), visceral adiposity index (VAI), and ultrasound measurements of subcutaneous and visceral adipose tissue depots and hepatic steatosis. WC and BMI showed the strongest correlations with ultrasound measurements of visceral adiposity. Only WHR correlated with sex hormones. Linear stepwise regression models including VAI were only slightly stronger than models including BMI or WC in explaining the variability in the insulin sensitivity index (yet BMI and WC had higher individual standardized coefficients of regression), and these models were superior to those including WHR and MOAD. WC showed 0.94 (95% confidence interval 0.88–0.99) and BMI showed 0.91 (0.85–0.98) probability of identifying the presence of hepatic steatosis according to receiver operating characteristic curve analysis. In conclusion, WC and BMI not only the simplest to obtain, but are also the most accurate surrogate markers of visceral adiposity in young adults, and are good indicators of insulin resistance and powerful predictors of the presence of hepatic steatosis.     

Structure and age jointly influence rates of protein evolution

What factors determine a protein's rate of evolution are actively debated. Especially unclear is the relative role of intrinsic factors of present-day proteins versus historical factors such as protein age. Here we study the interplay of structural properties and evolutionary age, as determinants of protein evolutionary rate. We use a large set of one-to-one orthologs between human and mouse proteins, with mapped PDB structures. We report that previously observed structural correlations also hold within each age group - including relationships between solvent accessibility, designabililty, and evolutionary rates. However, age also plays a crucial role: age modulates the relationship between solvent accessibility and rate. Additionally, younger proteins, despite being less designable, tend to evolve faster than older proteins. We show that previously reported relationships between age and rate cannot be explained by structural biases among age groups. Finally, we introduce a knowledge-based potential function to study the stability of proteins through large-scale computation. We find that older proteins are more stable for their native structure, and more robust to mutations, than younger ones. Our results underscore that several determinants, both intrinsic and historical, can interact to determine rates of protein evolution.     

Diversity and symbiotic effectiveness of <Emphasis Type="Italic">Adesmia</Emphasis> spp. root nodule bacteria in central and southern Chile

Legumes in the genus Adesmia are wild species with forage and medicinal potential. Their nitrogen fixation efficiency depends on their association with soil bacteria known as rhizobia. The aim of this work was to assess the diversity and symbiotic effectiveness of root nodule bacteria from Adesmia boronioides, Adesmia emarginata and Adesmia tenella from different regions of Chile. Adesmia spp. nodules were collected from seven sites obtaining 47 isolates, which resulted in 19 distinct strains. The diversity of the strains was determined via partial sequencing of the dnaK, 16srRNA and nodA genes. The strains were authenticated as root nodule bacteria on their original host and assessed for symbiotic effectiveness on A. emarginata and A. tenella. The strains from Adesmia tenella clustered within the Mesorhizobium clade. Adesmia boronioides nodulated with Mesorhizobium sp., Rhizobium leguminosarum and Bradyrhizobium sp. The rhizobia from A. emarginata were identified as Burkholderia spp, which was symbiotically ineffective on this species and on A. tenella. Strains isolated from Adesmia emarginata nodules, but unable to induce nodulation, were identified as Labrys methylaminiphilus. Labrys strain AG-49 significantly increased root dry weight in A. emarginata. The nodA genes from Adesmia strains were unique and correlated to legume host. A. emarginata was effectively nodulated by Bradyrhizobium AG-64 and A. tenella by Mesorhizobium strains AG-51 and AG.52. It is concluded that Adesmia emarginata, A. tenella and A. boronioides are associated to diverse bacterial symbionts and selection of an effective inoculant is a key step to assist Adesmia spp. adaptation and restoration.     

Effects of interleukin 17A (IL-17A) neutralization on murine hepatitis virus (MHV-A59) infection

Mice infected with mouse hepatitis virus A59 (MHV-A59) develop hepatitis and autoantibodies (autoAb) to liver and kidney fumarylacetoacetate hydrolase (FAH), a fact closely related to the release of alarmins such as uric acid and/or high-mobility group box protein 1 (HMGB1). We studied the effect of neutralizing monoclonal antibodies (MAb) against IL-17A in our model of mouse MHV-A59-infection. MAb anti-IL-17F and anti-IFNγ were used to complement the study. Results showed that transaminase levels markedly decreased in MHV-A59-infected mice treated with MAb anti-IL-17A whereas plasmatic Ig concentration sharply increased. Conversely, MAb anti-IL-17F enhanced transaminase liberation and did not affect Ig levels. Serum IFNγ was detected in mice infected with MHV-A59 and its concentration increased after MAb anti-IL-17A administration. Besides, MAb anti-IFNγ greatly augmented transaminase plasmatic levels. IL-17A neutralization did not affect MHV-A59-induction of HMGB1 liberation and slightly augmented plasmatic uric acid concentration. However, mice treated with the MAb failed to produce autoAb to FAH. The above results suggest a reciprocal regulation of Th1 and Th17 cells acting on the different MHV-A59 effects. In addition, it is proposed that IL-17A is involved in alarmins adjuvant effects leading to autoAb expression.