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King, L. J.

Publications and source records attributed to King, L. J..

2 recordsLinked to original sources

Background check: Mutational input to size variation depends on ancestor's breeding value

The phenotypic effects of mutations often depend on the genetic background, yet general patterns remain poorly resolved. Here, we tested whether genotypes drawn from the same natural population, but differing in their breeding values for a polygenic trait, differed in their contribution of new mutational variation to that trait. We established >200 mutation-accumulation (MA) lines from four Drosophila serrata genotypes. Analysing >44,000 wing-size measurements, collected over 30 generations, we quantified mutational variance and mutational bias for size. Genotypes with the smallest and largest breeding values for size contributed similar (statistically indistinguishable) amounts of mutational variance. In contrast, the genotype with an intermediate breeding value exhibited remarkably low (statistically undetectable) mutational variance, low micro-environmental variance, and high line survival over time, consistent with limited mutational decay in fitness. The three genotypes with detectable mutational input showed declines in mean size over time, indicating a consistent mutational bias toward smaller size, as reported in other taxa. The magnitude of this bias appeared genotype dependent, with the MA populations founded from the larger ancestors declining nearly twice as fast as that founded from the smallest ancestor. Together, these results demonstrate substantial heterogeneity in mutational properties among genotypes within a single natural population where the trait value spans a relatively narrow range. Such genotype-specific mutational input is expected to shape both the standing genetic variance and the evolutionary trajectory of polygenic traits.

evolutionary biology↗

Characterisation Of The Biflavonoid Hinokiflavone As A Pre-mRNA Splicing Modulator That Inhibits SENP

Here, we identify the plant biflavanoid hinokiflavone as an inhibitor of splicing in vitro and modulater of alternative splicing in multiple human cell lines. Hinokiflavone inhibits splicing in vitro by blocking one or more early steps of spliceosome assembly, leading to accumulation of the A complex. Multiple human cell lines treated with hinokiflavone show changes in the alternative splicing of different pre-mRNA substrates, but little or no change in transcription. They also show altered subnuclear organization, specifically of splicing factors required for A complex formation, which relocalized together with SUMO1 and SUMO2 into enlarged nuclear speckles. While most cell lines treated with hinokiflavone showed cell cycle arrest and eventual cell death, dependent on time and concentration, the promyelocytic NB4 cell line, which expresses the SUMO target PML-RARalpha fusion protein, was exquisitely sensitive to apoptosis following hinokiflavone treatment. Hinokiflavone treatment increased protein SUMOylation levels, both in in vitro splicing reactions and in cells, with little or no effect on levels of ubiquitinylated proteins. Hinokiflavone also inhibited the catalytic activity of purified E. coli expressed SUMO protease, SENP1 in vitro, indicating the increase in SUMOylated proteins results primarily from inhibition of de-SUMOylation. Using a quantitative proteomics assay we identified many SUMO2 sites whose levels increased following hinokiflavone treatment, with the major targets including 6 proteins that are associated with U2 snRNP and required for A complex formation. These data identify hinokiflavone as a SUMO protease inhibitor and indicate SUMOylation of splicing factors may be important for modulating splice site selection.

biochemistry↗