Ablating putative Ku70 phosphorylation sites results in defective DNA damage repair and spontaneous induction of hepatocellular carcinoma.

Multiple pathways mediate the repair of DNA double-strand breaks (DSBs), with numerous mechanisms responsible for driving choice between the pathways. Previously, we reported that mutating five putative phosphorylation sites on the non-homologous end joining (NHEJ) factor, Ku70, results in sustained retention of human Ku70/80 at DSB ends and attenuation of DSB repair via homologous recombination (HR). In this study, we generated a knock-in mouse, in which the three conserved putative phosphorylation sites of Ku70 were mutated to alanine to ablate potential phosphorylation (Ku703A/3A), in order to examine if disrupting DSB repair pathway choice by modulating Ku70/80 dynamics at DSB ends results in enhanced genomic instability and tumorigenesis. The Ku703A/3A mice developed spontaneous and have accelerated chemical-induced hepatocellular carcinoma (HCC) compared to wild-type (Ku70+/+) littermates. The HCC tumors from the Ku703A/3A mice have increased γH2AX and 8-oxo-G staining, suggesting decreased DNA repair. Spontaneous transformed cell lines from Ku703A/3A mice are more radiosensitive, have a significant decrease in DNA end resection, and are more sensitive to the DNA cross-linking agent mitomycin C compared to cells from Ku70+/+ littermates. Collectively, these findings demonstrate that mutating the putative Ku70 phosphorylation sites results in defective DNA damage repair and disruption of this process drives genomic instability and accelerated development of HCC.

Nitrate repletion strategy for enhancing lipid production from marine microalga Tetraselmis sp.

The cell growth rate and cellular lipid content of microalgae are affected by the nitrogen levels during cultivation. The growth rate and lipid content of marine microalga Tetraselmis sp. was found to increase under nitrate replete conditions, but not under deplete conditions. Thus, in order to enhance the lipid productivity of Tetraselmis sp., a two-stage culture process utilizing nitrate replete condition was applied. When the cells were cultivated in F/2 medium for five days in the first stage, the obtained lipid content and productivity were 22.4% and 26.7mgL(-1)d(-1), respectively. After second stage of cultivation for a further 36h under nitrate replete conditions with 8.82mM NaNO3, increased biomass concentration of 1.32gL(-1) and lipid content of 30.5% were obtained, with an enhanced lipid productivity of 47.3mgL(-1)d(-1).