Genetic differentiation and phylogenetic potential of Ty3/Gypsy LTR retrotransposon markers in soil and plant pathogenic fungi

Genetic diversity studies are crucial for understanding the genetic structure and evolutionary dynamics of fungal species and communities. Fungal genomes are often reshaped by their repetitive components such as transposable elements. These elements are key players in genomic rearrangements and are ideal targets for genetic diversity and evolutionary studies. Herein, we used three Ty3/Gypsy long terminal repeat retrotransposons, Grasshopper, Maggy, and Pyret, for genetic differentiation and diversity in soil and plant pathogenic fungi, representing diverse species, order, and phyla. Pyret DNA markers showed the highest gene diversity and Shannon's information indices, followed by Maggy and Grasshopper. The observed high levels of multilocus polymorphism indicate the continuous mobility of these elements after their transfer in the new host. In conclusion, this study presents novel markers for genetic differentiation and evolutionary studies of fungi, and sheds light on the prevalence of gene acquisition phenomenon in field fungi.     

Adaptive response in stress granule formation and bulk translational repression upon a combined stress of mild heat shock and mild ethanol stress in yeast

The formation of cytoplasmic mRNA–protein complex granules termed ‘processing bodies and stress granules’ is often induced in the stress responses of eukaryotic cells. Most previous studies on stress granules have focused on the response to a single type of stress, and little information is available about the response to combined stress. Additionally, the effects of adaptation on stress granule formation and bulk translation activity are poorly understood. We investigated the formation of stress granules upon combined exposure to mild heat shock (37 °C) and mild ethanol stress (5% v/v) in Saccharomyces cerevisiae. Although neither stress alone induced stress granule formation, their combination caused a pronounced repression of translation activity and the formation of stress granules. Pretreatment with each mild stress significantly attenuated the formation of stress granules and caused changes in the composition of stress granules upon the subsequent combined stress and facilitated stress granule disassembly accompanied by smooth translational resurrection during the recovery process, indicating that yeast cells can induce adaptations in stress granule formation. However, the pretreated cells still exhibited a severe repression of translation activity. These findings provide novel and fundamental insight into the regulation of yeast stress granules.     

Genome-wide analysis of cellular response to bacterial genotoxin CdtB in yeast

The cytolethal distending toxins (CDTs) are secreted virulence proteins produced by several bacterial pathogens, and the subunit CdtB has the ability to create DNA lesions, primarily DNA single-strand breaks (SSBs) in vitro, and cause cell cycle arrest, cellular distension, and cell death in both mammalian and yeast cells. To elucidate the components of the mechanisms underlying the response to CdtB-induced DNA lesions, a CdtB expression plasmid was transformed into a series of diploid yeast strains harboring deletions in 4,708 nonessential genes. A total of 4,706 of these clones were successfully transformed, which we have now designated as a systematic transformation array (STA), and were subsequently screened. We identified 61 sensitive strains from the STA whose deleted genes can be categorized into a number of groups, including DNA metabolism, chromosome segregation, vesicular traffic, RNA catabolism, protein translation, morphogenesis, and nuclear transport, as well as one unknown open reading frame. However, only 28 of these strains were found to be sensitive to HO endonuclease, which is known to create a DNA double-strand break (DSB), suggesting that CdtB-induced DNA lesion is not similar to the direct DSB. Amazingly, CdtB expression elicits severe growth defects in haploid yeast cells, but only marginal defects in diploid yeast cells. The presence and absence of genes known to be involved in DNA repair in these genome-wide data reveal that CdtB-induced DNA damage is specifically repaired well in the diploid by homologous recombination but not by other repair mechanisms. Our present results provide insights into how CdtB pathogenesis is linked to eukaryotic cellular functions.     

Age differences in the body temperature response to restraint-cold stress

Female Fischer 344 rats, 3, 11, and 24-months old were subjected to 3 hr of restraint-cold stress once each week for 4 weeks. Body (colon) temperature was recorded every 10 min during restraint and during postrestraint recovery until temperature returned to the prerestraint level. Thermoregulatory adaptation during consecutive restraint sessions was noted in 3- and 11-month-old rats but not in 24-month-old rats. Significant temperature recovery was accomplished after the second restraint session for 3- and 11-month-old rats, but this was not accomplished in the older 24-month rats until after four restraint sessions. The role of hypothermia in restraint-induced ulceration is questioned, especially as it pertains to elderly rats.     

Sex-associated differences in the antibody-dependent cellular cytotoxicity antibody response to measles vaccines

In some countries, excessive non-measles-related mortality has been observed among female recipients of high-titer measles vaccines. We determined if differences in the immune response to measles vaccines underlie the excessive female mortality by measuring the measles virus (MV)-specific antibody-dependent cellular cytotoxicity (ADCC) antibody response in 65 3-year-old Gambian children immunized with Edmonston-Zagreb medium-titer (EZ) or Schwarz standard vaccines during infancy. Among the 20 females and 22 males with undetectable anti-MV antibodies at the time of immunization, females had significantly lower ADCC than males (median cytotoxicities of 1/100 serum dilutions = 8.4 and 12%, respectively; P = 0.04). This sex-associated difference was present only among the six female and seven male recipients of EZ vaccine (median cytotoxicities = 5.1 and 19.0%, respectively; P = 0.02). There were no significant sex-associated differences in neutralizing antibody activity. Decreased ADCC antibody activity may contribute to the lower survival rate observed in females receiving high-titer measles vaccination.     

Sex differences in the interleukin-6 response to acute psychological stress

Interleukin-6 (IL-6), an immune regulator that helps coordinate the inflammatory response, may mediate inflammatory disease exacerbation associated with stress. Twenty men and twenty women completed a single session, comprising baseline (20 min), mental arithmetic task (8 min), and recovery (60 min). Blood samples, taken at baseline, immediately after the task, and at +30 and +60 min recovery were analysed for plasma IL-6. Overall, IL-6 increased linearly from baseline to +60 min recovery, and a sex difference was found in the IL-6 response, with men peaking earlier than women. These findings confirm a small delayed IL-6 increase after acute laboratory stress, and reveal sex differences in the profile of the IL-6 response.     

ATM is required for the cellular response to thymidine induced replication fork stress

Genetically distinct checkpoints, activated as a consequence of either DNA replication arrest or ionizing radiation-induced DNA damage, integrate DNA repair responses into the cell cycle programme. The ataxia-telangiectasia mutated (ATM) protein kinase blocks cell cycle progression in response to DNA double strand breaks, whereas the related ATR is important in maintaining the integrity of the DNA replication apparatus. Here, we show that thymidine, which slows the progression of replication forks by depleting cellular pools of dCTP, induces a novel DNA damage response that, uniquely, depends on both ATM and ATR. Thymidine induces ATM-mediated phosphorylation of Chk2 and NBS1 and an ATM-independent phosphorylation of Chk1 and SMC1. AT cells exposed to thymidine showed decreased viability and failed to induce homologous recombination repair (HRR). Taken together, our results implicate ATM in the HRR-mediated rescue of replication forks impaired by thymidine treatment.     

RpoS differentially affects the general stress response and biofilm formation in the endophytic Serratia plymuthica G3

The σ^S subunit RpoS of RNA polymerase functions as a master regulator of the general stress response in Escherichia coli and related bacteria. RpoS has been reported to modulate biocontrol properties in the rhizobacterium Serratia plymuthica IC1270. However, the role of RpoS in the stress response and biofilm formation in S. plymuthica remains largely unknown. Here we studied the role of RpoS from an endophytic S. plymuthica G3 in regulating these phenotypes. Mutational analysis demonstrated that RpoS positively regulates the global stress response to acid or alkaline stresses, oxidative stress, hyperosmolarity, heat shock and carbon starvation, in addition to proteolytic and chitinolytic activities. Interestingly, rpoS mutations resulted in significantly enhanced swimming motility, biofilm formation and production of the plant auxin indole-3-acetic acid (IAA), which may contribute to competitive colonization and environmental fitness for survival. These findings provide further insight into the strain-specific role of RpoS in the endophytic strain G3 of S. plymuthica, where it confers resistance to general stresses encountered within the plant environment. The heterogeneous functionality of RpoS in rhizosphere and endophytic S. plymuthica populations may provide a selective advantage for better adaptation to various physiological and environmental stresses.     

Tagging chromatin with retrotransposons: target specificity of the Saccharomyces Ty5 retrotransposon changes with the chromosomal localization of Sir3p and Sir4p

Retrotransposon and retroviral insertions are not randomly distributed on chromosomes, suggesting that retroelements actively select integration sites. This is the case for the yeast Ty5 retrotransposons, which preferentially integrate into domains of silent chromatin at the HM loci and telomeres. Here we demonstrate that loss of Sir3p or Sir4p-components of silent chromatin-causes a greater than ninefold decrease in Ty5 targeting to the HM loci and largely randomizes chromosomal integration patterns. Strains with a deletion of SIR4 also display an approximately 10-fold increase in cDNA recombination, which is due both to the expression a- and alpha-mating-type information and the loss of Sir4p. It is known that in old yeast cells or in strains carrying the sir4-42 allele, the Sir complex relocalizes to the rDNA. About 26% of Ty5 insertions occur within the rDNA in sir4-42 strains compared with 3% in wild type. Ty5, therefore, is sensitive to changes in chromatin, indicating that retrotransposons may be useful for dissecting chromatin dynamics that occur during developmental programs such as aging.     

Age differences in the hormonal response of the adrenals and testes to stress in monkeys

Adrenal and testicular function of sexually immature and adult male baboons (Papio hamadryas) was investigated during immobilization stress. The concentrations of testosterone, 5-dihydrotestosterone, hydrocortisone, and intermediate products of their biosynthesis — progesterone, pregnenolone, 17-hydroxyprogesterone, 17-hydroxypregnenolone, and 11-deoxycortisol — in the peripheral blood plasma of the monkeys were determined by a radioimmunological method. The main age differences in the character of the hormonal response to stress were shown to be the absence of changes in the blood androgen concentration of the immature animals and the smaller increase in the hydrocortisone concentration in them than in the adult monkeys. The concentrations of precursors of the steroid hormones fell considerably in the immature monkeys during stress, whereas in the adult animals their level rose 3–4-fold.Laboratory of Biochemistry, Institute of Experimental Pathology and Therapy, Academy of Medical Sciences of the USSR, Sukhumi. (Presented by Academician of the Academy of Medical Sciences of the USSR B. A. Lapin.) Translated from Byulleten' Éksperimental'noi Biologii i Meditsiny, Vol. 89, No. 2, pp. 157–160, February, 1980.     

Regulatory role of Epo3’-enhancer in cellular hypoxic response

Regulatory role of Epo3’-enhancer in cellular hypoxic response     

Sex differences in sleep: the response to sleep deprivation and restraint stress in mice

STUDY OBJECTIVES: Numerous clinical studies and sleep surveys have shown pronounced sex differences in the occurrence of insomnia and other sleep pathologies. It has been suggested that sex differences in sleep, while subtle under baseline conditions, may increase in magnitude under biological or environmental challenges. However, controlled and experimental studies on sleep under challenged conditions rarely include female subjects. In this context, we examined sex differences in sleep in the mouse, not only under baseline conditions, but also after sleep deprivation and restraint stress. DESIGN: Adult male and female C57BL/6J mice were implanted with electrodes to record sleep-wake architecture and sleep electroencephalogram under baseline conditions and after 6 hours of sleep deprivation or 1 hour of restraint stress at the beginning of the daily light phase. RESULTS: Although baseline sleep patterns slightly differed between the sexes, the homeostatic recovery response to sleep deprivation was similar. In contrast, the changes in sleep after restraint stress were markedly different between male and female mice, with males displaying a stronger initial suppression of sleep and a stronger rebound of rapid-eye-movement sleep later in the recovery phase. CONCLUSIONS: In mice, the fundamental homeostatic properties of sleep regulation may not differ between the sexes, but the way sleep is affected and disrupted by environmental influences may be sex dependent. The latter may reflect sex differences in stress sensitivity.     

Relationships between the epiphysis and hippocampus during formation of a stress response

Lesions to the dorsal hippocampus and removal of the epiphysis had differently-directed effects on behavior in rats in conditions of a conflict situation and on the temporal dynamics of forced swimming. The combination of both operative procedures resulted in weakening of the behavioral changes typical of hippocampus deficiency. It is suggested that the anti-stress properties of epiphyseal factors may be mediated by changes in the functional state of the hippocampus. The Medical Academy, Stavropol'. Translated from Zhurnal Vysshei Nervnoi Deyatel'nosti imeni I. P. Pavlova, Vol. 47, No. 4, pp. 725–730, July–August, 1997.