Watching right and wrong nucleotide insertion captures hidden polymerase fidelity checkpoints.

Efficient and accurate DNA synthesis is enabled by DNA polymerase fidelity checkpoints that promote insertion of the right instead of wrong nucleotide. Erroneous X-family polymerase (pol) λ nucleotide insertion leads to genomic instability in double strand break and base-excision repair. Here, time-lapse crystallography captures intermediate catalytic states of pol λ undergoing right and wrong natural nucleotide insertion.

Structural basis for proficient oxidized ribonucleotide insertion in double strand break repair.

Reactive oxygen species (ROS) oxidize cellular nucleotide pools and cause double strand breaks (DSBs). Non-homologous end-joining (NHEJ) attaches broken chromosomal ends together in mammalian cells. Ribonucleotide insertion by DNA polymerase (pol) μ prepares breaks for end-joining and this is required for successful NHEJ in vivo.

Watching a double strand break repair polymerase insert a pro-mutagenic oxidized nucleotide.

Oxidized dGTP (8-oxo-7,8-dihydro-2´-deoxyguanosine triphosphate, 8-oxodGTP) insertion by DNA polymerases strongly promotes cancer and human disease. How DNA polymerases discriminate against oxidized and undamaged nucleotides, especially in error-prone double strand break (DSB) repair, is poorly understood. High-resolution time-lapse X-ray crystallography snapshots of DSB repair polymerase μ undergoing DNA synthesis reveal that a third active site metal promotes insertion of oxidized and undamaged dGTP in the canonical anti-conformation opposite template cytosine.

Preferential DNA Polymerase β Reverse Reaction with Imidodiphosphate.

DNA replication and repair reactions involve the addition of a deoxynucleoside monophosphate onto a growing DNA strand with the loss of pyrophosphate. This chemical reaction is also reversible; the addition of pyrophosphate generates a deoxynucleoside triphosphate, thereby shortening the DNA by one nucleotide. The forward DNA synthesis and reverse pyrophosphorolysis reactions strictly require the presence of divalent metals, usually magnesium, at the reactive center as cofactors.

The economic impact that registered veterinary technicians have on Ontario veterinary practices.

In Canada, registered veterinary technicians (RVTs) are highly trained, regulated professionals working as an integral part of the veterinary team, but there has been little work to quantify their impact on veterinary practices. The objectives of the study were to explore the utilization rates of RVTs and quantify the economic value that RVTs bring to veterinary practices in Ontario, Canada. An online survey was deployed to explore the function of RVTs and their non-credentialed counterparts in practices.