Biology subjects

Humphreys, J. M.

Publications and source records attributed to Humphreys, J. M..

3 recordsLinked to original sources

bioRxiv2025

Spatial transmission dynamics of African Swine Fever in wild pigs: Sensitivity to epidemiological traits of different viral strains

African Swine Fever virus (ASFv) is a rapidly spreading animal disease with impacts on global economy and food security. Since the first known case of ASFv in Kenya in 1921, the virus has undergone significant evolution resulting in the emergence of 25 distinct genotypes. These genotypes have a wide variety of epidemiological traits, leading to uncertainty about how a genotype or variant of a genotype might spread in a population. Understanding the impacts of different epidemiological traits on the spread of ASFv is important for developing effective surveillance systems and control strategies. We developed a spatial epidemiological model of ASFv transmission in wild pigs, which contributes to ASFv persistence in countries where wild pig populations are widespread. We evaluated the impacts of a realistic set of epidemiological traits including incubation period, infectious period, disease-induced mortality rate, and length of immunity on spatial transmission dynamics under different ecological conditions for the host population (i.e. host density). We analyze effects of these conditions on outbreak metrics using boosted regression models with response variables: peak incidence, rate of spatial spread, seroprevalence and area invaded after one year, and probability that an outbreak will occur. We found that the infectious period of living individuals is the most important predictor at all pig densities for peak incidence and the area affected at one year, followed closely by the incubation period of the virus. We discuss considerations for surveillance and control strategies for different genotypes of ASFv.

ecology↗

bioRxiv2023

Water and chloride as allosteric inhibitors in WNK kinase osmosensing

Osmotic stress and chloride regulate the autophosphorylation and activity of the WNK1 and WNK3 kinase domains. The kinase domain of unphosphorylated WNK1 (uWNK1) is an asymmetric dimer possessing water molecules conserved in multiple uWNK1 crystal structures. Conserved waters are present in two networks, referred to here as conserved water networks 1 and 2 (CWN1and CWN2). Here we show that PEG400 applied to crystals of dimeric uWNK1 induces de-dimerization. Both the WNK1 the water networks and the chloride binding site and are disrupted by PEG400. CWN1 is surrounded by a cluster of pan-WNK-conserved charged residues. Here we mutagenized these charges in WNK3, a highly active WNK isoform kinase domain, and WNK1, the isoform best studied crystallographically. Mutation of E314 in the Activation Loop of WNK3 (WNK3/E314Q and WNK3/E314A, and the homologous WNK1/E388A) enhanced the rate of autophosphorylation, and reduced chloride sensitivity. Other WNK3 Cluster mutants reduced the rate of autophosphorylation activity coupled with greater chloride sensitivity than wild-type. The water and chloride regulation thus appear linked. The lower activity of some mutants may reflect effects on catalysis. Crystallography showed that activating mutants introduced conformational changes in similar parts of the structure to those induced by PEG400. WNK activating mutations and crystallography support a role for CWN1 in WNK inhibition consistent with water functioning as an allosteric ligand.

biophysics↗

bioRxiv2018

Anatomy of a pressure sensing protein kinase

Cells respond to hydrostatic pressure to maintain cellular, organ, and organism level functions, yet the direct pressure sensors are largely unknown. Here we show that hydrostatic pressure directly activates With No Lysine(K) kinase-3 (WNK3) 1, a soluble intracellular protein kinase. Using gel filtration we demonstrate that pressure induces a dimer to monomer transition in a construct of the unphosphorylated kinase domain of WNK3 (uWNK3-KDm or uWNK3). The uWNK3 has not been crystallized, but crosslinking data suggest that the uWNK3 dimer corresponds to crystallographically observed dimer of WNK1 (uWNK1-KDm, or uWNK1) 2,3. Sequence alignments with WNKs from species living in different pressure environments and mutational analysis lend further support for this idea. Unique features of the uWNK1 structure suggest a mechanism involving bound water. We further show that hydrostatic pressure activates full-length WNK3 in D. melanogaster tubules.

biophysics↗