The Deletion of the Bre1 Gene in Aspergillus nidulans Impairs Mitotic Growth, Meiosis, and DNA Damage Repair

Bre1 is a homotetrameric E3 ubiquitin-protein ligase that heterodimerizes with Rad6, an E2 ubiquitin-conjugating enzyme, in order to ubiquitinate lysine 123 in Aspergillus nidulans. This post-translational modification promotes methylation of lysines 4 and 79 on histone H3, which are required for certain damage repair pathways and for both optimal mitotic cell growth and meiosis [1-3, 12]. ΔBre1 mutants were generated by exposing protoplasts from strains auxotrophic for pyridoxine to a three-way fusion construct made from the Bre1 5’ and 3’ flanking regions and the Aspergillus fumigatus pyroA gene, which served as a selectable marker. Molecular diagnosis was confirmed via trans-locus PCR. Phenotypic analysis indicates that the loss of Bre1 increases sensitivity to DNA damage agents, decreases mitotic cell growth, and inhibits meiosis. The severe developmental defects of ΔBre1 mutants are consistent with the known roles of Bre1 as an upstream regulator of several important cellular functions. [excerpt

Differential Uptake of Gold Nanoparticles by 2 Species of Tadpole, the Wood Frog (Lithobates Sylvaticus) and the Bullfrog (Lithobates Catesbeianus)

Engineered nanoparticles are aquatic contaminants of emerging concern that exert ecotoxicological effects on a wide variety of organisms. We exposed cetyltrimethylammonium bromide–capped spherical gold nanoparticles to wood frog and bullfrog tadpoles with conspecifics and in combination with the other species continuously for 21 d, then measured uptake and localization of gold. Wood frog tadpoles alone and in combination with bullfrog tadpoles took up significantly more gold than bullfrogs. Bullfrog tadpoles in combination with wood frogs took up significantly more gold than controls. The rank order of weight-normalized gold uptake was wood frogs in combination \u3e wood frogs alone \u3e bullfrogs in combination \u3e bullfrogs alone \u3e controls. In all gold-exposed groups of tadpoles, gold was concentrated in the anterior region compared with the posterior region of the body. The concentration of gold nanoparticles in the anterior region of wood frogs both alone and in combination with bullfrogs was significantly higher than the corresponding posterior regions. We also measured depuration time of gold in wood frogs. After 21 d in a solution of gold nanoparticles, tadpoles lost \u3e83% of internalized gold when placed in gold-free water for 5 d. After 10 d in gold-free water, tadpoles lost 94% of their gold. After 15 d, gold concentrations were below the level of detection. Our finding of differential uptake between closely related species living in similar habitats with overlapping geographical distributions argues against generalizing toxicological effects of nanoparticles for a large group of organisms based on measurements in only one species

Long-term Exposure to Gold Nanoparticles Accelerates Larval Metamorphosis Without Affecting Mass in Wood Frogs (Lithobates sylvaticus) at Environmentally Relevant Concentrations

Nanoparticles are environmental contaminants of emerging concern. Exposure to engineered nanoparticles has been shown to have detrimental effects on aquatic organisms. We synthesized gold nanoparticles (18.1 ± 3.5 nm) and tested their effects on time to and weight at metamorphosis in wood frog (Lithobates sylvaticus) tadpoles, a species known to be sensitive to environmental stressors. Continuous exposure to all concentrations of gold nanoparticles (0.05, 0.5, and 5 pM in particles) for up to 55 days significantly reduced time to metamorphosis by as much as an average of three days (p \u3c 0.05). However, exposure to gold nanoparticles had no effect on tadpole mass at metamorphosis. The ∼18 nm gold nanoparticles used in this study were metastable in dechlorinated tap water resulting in a change in surface charge and aggregation over time leading to negatively charged aggregates that are on the order of 60–110 nm. Nanoparticle aggregation could exacerbate the effect on time to metamorphosis. To our knowledge, this is the first report on the effect of engineered nanoparticles of any kind on life history variables in an amphibian, a taxonomic group that has been declining globally for at least 25 years