Effects of diesel exhaust on the microbiota within a tuffaceous tunnel system

The abundance and distribution of microbiota that may be impacted by diesel and diesel exhaust were investigated from three depths into the walls and invert (floor) of U12n tunnel at Rainier Mesa, Nevada Test Site, a potential geological analog of Yucca Mountain. Enumerations included total cell counts, and numbers of aerobic heterotrophic, sulfate-reducing, nitrate-reducing, and diesel-degrading bacteria. Additionally, the disappearance of total petroleum hydrocarbons was determined in microcosms containing subsurface materials that were amended with diesel fuel. Results revealed that microbes capable of utilizing diesel and diesel combustion products were present in the subsurface in both the walls and the invert of the tunnel. The abundance of specific bacterial types in the tunnel invert, a perturbed environment, was greater than that observed in the tunnel wall. Few trends of microbial distribution either into the tunnel wall or the invert were noted with the exception of aerobic heterotrophic abundance which increased with depth into the wall and decreased with depth into the invert. No correlation between microbiota and a specific introduced chemical species have yet been determined. The potential for microbial contamination of the tunnel wall during sampling was determined to be negligible by the use of fluorescently labeled latex spheres (1{mu}m in dia.) as tracers. Results indicate that additional investigations might be needed to examine the microbiota and their possible impacts on the geology and geochemistry of the subsurface, both indigenous microbiota and those microorganisms that will likely be introduced by anthropogenic activity associated with the construction of a high-level waste repository

An E2F1-Mediated DNA Damage Response Contributes to the Replication of Human Cytomegalovirus

DNA damage resulting from intrinsic or extrinsic sources activates DNA damage responses (DDRs) centered on protein kinase signaling cascades. The usual consequences of inducing DDRs include the activation of cell cycle checkpoints together with repair of the damaged DNA or induction of apoptosis. Many DNA viruses elicit host DDRs during infection and some viruses require the DDR for efficient replication. However, the mechanism by which DDRs are activated by viral infection is poorly understood. Human cytomegalovirus (HCMV) infection induces a DDR centered on the activation of ataxia telangiectasia mutated (ATM) protein kinase. Here we show that HCMV replication is compromised in cells with inactivated or depleted ATM and that ATM is essential for the host DDR early during infection. Likewise, a downstream target of ATM phosphorylation, H2AX, also contributes to viral replication. The ATM-dependent DDR is detected as discrete, nuclear γH2AX foci early in infection and can be activated by IE proteins. By 24 hpi, γH2AX is observed primarily in HCMV DNA replication compartments. We identified a role for the E2F1 transcription factor in mediating this DDR and viral replication. E2F1, but not E2F2 or E2F3, promotes the accumulation of γH2AX during HCMV infection or IE protein expression. Moreover, E2F1 expression, but not the expression of E2F2 or E2F3, is required for efficient HCMV replication. These results reveal a novel role for E2F1 in mediating an ATM-dependent DDR that contributes to viral replication. Given that E2F activity is often deregulated by infection with DNA viruses, these observations raise the possibility that an E2F1-mediated mechanism of DDR activation may be conserved among DNA viruses