Ku controls telomere replication timing. We test the mechanism and find that Ku does not bind telomere-proximal origins directly or alter their histone acetylation state. Instead, Ku's effect on replication timing is mediated through telomere length and requires the TG1-3 repeat-counting component Rif1.

Ure2 is the protein determinant of the [URE3] prion phenotype inHere we tested the effect of overexpression of Hsp40 members Ydj1, Sis1, and Apj1 and also Hsp70 co-chaperones Cpr7, Cns1, Sti1, and Fes1 in vivo and found that only Ydj1 showed a strong curing effect on [URE3]. We also investigated the interaction of Ydj1 with Ure2 in vitro. We found that Ydj1 was able to suppress formation of amyloid-like fibrils of Ure2 by delaying the process of fibril formation, as monitored by thioflavin T binding and atomic force microscopy imaging. Controls using bovine serum albumin, Sis1, or the human Hsp40 homologues Hdj1 or Hdj2 showed no significant inhibitory effect. Ydj1 was only effective when added during the lag phase of fibril formation, suggesting that it interacts with Ure2 at an early stage in fibril formation and delays the nucleation process. Using surface plasmon resonance and size exclusion chromatography, we demonstrated a direct interaction between Ydj1 and both wild type and N-terminally truncated Ure2. In contrast, Hdj2, which did not suppress fibril formation, did not show this interaction. The results suggest that Ydj1 inhibits Ure2 fibril formation by binding to the native state of Ure2, thus delaying the onset of oligomerization.The epigenetic factor [URE3] in the yeast Saccharomyces cerevisiae represents the prion form of the protein Ure2 (1). Like the mammalian prion, the heritable [URE3] phenotype is conveyed by a structural change in its protein determinant to an aggregated form (1, 2). Ure2 is a 354-amino acid homodimeric protein consisting of a relatively flexible and protease-sensitive N-terminal region (ϳ90 amino acids) and a globular C-terminal region (3-5). The N-terminal region is required for its prion properties in vivo (2) and to form amyloid-like filaments in vitro (6 -8). However, deletion of the N-terminal region has no detectable effect on the stability or folding of the protein in vitro (9). The C-terminal region, for which the crystal structure has been determined in both apo (5, 10) and glutathione-bound (11) forms, shows structural similarity to glutathione transferases (GSTs), 3 and is necessary and sufficient for its regulatory function in vivo: Ure2 interacts with the transcription factor Gln3, allowing control of nitrogen catabolite repression and blocking the uptake of poor nitrogen sources in the presence of a good nitrogen source (12, 13). In addition, Ure2 possesses glutathione-dependent peroxidase (GPx) activity, which is maintained upon formation of fibrillar aggregates, indicating that the C-terminal globular domains of Ure2 retain their native structure within the fibrils (14).Ydj1 from S. cerevisiae is a molecular chaperone of the type I Hsp40 family and is involved in multiple functions, including import of proteins into mitochondria, secretion of mating pheromones, and regulation of the activity of the cytoplasmic Hsp70s (15-17). Ydj1 can bind to nonnative polypeptides and pair with Hsp70 Ssa proteins to prevent aggregation and facilitate refolding of denatured p...

The Saccharomyces cerevisiae protein Ure2 functions as a regulator of nitrogen metabolism and as a glutathione-dependent peroxidase. Ure2 also has the characteristics of a prion, in that it can undergo a heritable conformational change to an aggregated state; the prion form of Ure2 loses the regulatory function, but the enzymatic function appears to be maintained. A number of factors are found to affect the prion properties of Ure2, including mutation and expression levels of molecular chaperones, and the effect of these factors on structure and stability are being investigated. The relationship between structure, function and folding for the yeast prion Ure2 are discussed.

No abstract

A literal mountain of documentation generated in the past five decades showing unmistakable health hazards associated with extremely low-frequency electromagnetic fields (ELF-EMFs) exposure. However, the relation between energy mechanism and ELF-EMF exposure is poorly understood. In this study, Caenorhabditis elegans was exposed to 50 Hz ELF-EMF at intensities of 0.5, 1, 2, and 3 mT, respectively. Their metabolite variations were analyzed by GC-TOF/MS-based metabolomics. Although minimal metabolic variations and no regular pattern were observed, the contents of energy metabolism-related metabolites such as pyruvic acid, fumaric acid, and L-malic acid were elevated in all the treatments. The expressions of nineteen related genes that encode glycolytic enzymes were analyzed by using quantitative real-time PCR. Only genes encoding GAPDH were significantly upregulated (P < 0.01), and this result was further confirmed by western blot analysis. The enzyme activity of GAPDH was increased (P < 0.01), whereas the total intracellular ATP level was decreased. While no significant difference in lifespan, hatching rate and reproduction, worms exposed to ELF-EMF exhibited less food consumption compared with that of the control (P < 0.01). In conclusion, C. elegans exposed to ELF-EMF have enhanced energy metabolism and restricted dietary, which might contribute to the resistance against exogenous ELF-EMF stress.

Cancer, as a leading cause of death, has attracted enormous public attention. Reprogramming of cellular energy metabolism is deemed to be one of the principal hallmarks of cancer. In this article, we reviewed the mutual relationships among environmental pollution factors, energy metabolic dysfunction, and various cancers. We found that most environmental pollution factors could induce cancers mainly by disturbing the energy metabolism. By triggering microenvironment alteration, energy metabolic dysfunction can be treated as a factor in carcinogenesis. Thus, we put forward that energy metabolism might be as a key point for studying carcinogenesis and tumor development to propose new methods for cancer prevention and therapy.

Temperature is an important factor in research on the biological effects of extremely low-frequency electromagnetic field (ELF-EMF), but interactions between ELF-EMF and temperature remain unknown. The effects of ELF-EMF (50 Hz, 3 mT) on the lifespan, locomotion, heat shock response (HSR), and oxidative stress (OS) of Canton-Special (CS) and mutant w1118 flies were investigated at 25°C and 35°C (thermal stress). Results showed that thermal stress accelerated the death rates of CS and w1118 flies, shortened their lifespan, and influenced their locomotion rhythm and activity. The upregulated expression levels of heat shock protein (HSP) 22, HSP26, and HSP70 indicated that HSR was enhanced. Thermal stress-induced OS response increased malondialdehyde content, enhanced superoxide dismutase activity, and decreased reactive oxygen species level. The effects of thermal stress on the death rates, lifespan, locomotion, and HSP gene expression of flies, especially w1118 line, were also enhanced by ELF-EMF. In conclusion, thermal stress weakened the physiological function and promoted the HSR and OS of flies. ELF-EMF aggravated damages and enhanced thermal stress-induced HSP and OS response. Therefore, thermal stress and ELF-EMF elicited a synergistic effect.

In this study, we studied the effect of 2.0 GHz radio frequency electromagnetic field (RF-EMF) and 50 Hz extremely low frequency electromagnetic field (ELF-EMF) exposure on prion generation and propagation using two budding yeast strains, NT64C and SB34, as model organisms. Under exposure to RF-EMF or ELF-EMF, the de novo generation and propagation of yeast prions [URE3] were elevated in both strains. The elevation increased over time, and the effects of ELF-EMF occurred in a dose-dependent manner. The transcription and expression levels of the molecular chaperones Hsp104, Hsp70-Ssa1/2, and Hsp40-Ydj1 were not statistically significantly changed after exposure. Furthermore, the levels of ROS, as well as the activities of superoxide dismutase (SOD) and catalase (CAT), were significantly elevated after short-term, but not long-term exposure. This work demonstrated for the first time that EMF exposure could elevate the de novo generation and propagation of yeast prions and supports the hypothesis that ROS may play a role in the effects of EMF on protein misfolding. The effects of EMF on protein folding and ROS levels may mediate the broad effects of EMF on cell function.