Search bioRxiv⌕ Search

Biology subjects

Laurent, J.

Publications and source records attributed to Laurent, J..

3 recordsLinked to original sources

Medium-assisted tumbling controls bacteria exploration in a complex fluid

In nature, many fluids that harbor bacterial populations or protect against microbial contamination exhibit non-Newtonian rheology. To study the spatial exploration of E.coli bacteria, a model multi-flagellated microorganism, in such complex environments, we design a motility medium with tunable macroscopic rheology. By increasing the solid charge in soft carbomer grains, we transition from a Newtonian viscous suspension to a yield-stress fluid. Using a 3D Lagrangian tracking device, we collect many individual bacterial tracks and characterize changes in motility properties such as swimming speed, persistence times and diffusivity for both a wild-type and a smooth runner mutant, up to the formation of a motility barrier at higher carbomer concentrations. We show that the presence of local mechanical disorder and resistance to penetration essentially override the biologically driven run-and-tumble navigation process. This "medium-assisted" exploration scenario is characterized by directional switching and stop-and-go kinematics and is closely related to the flexibility of the flagellar bundle.

biophysics↗

ACLY promotes NK cell effector function by regulating glycolysis and histone acetylation

Natural Killer (NK) cells are innate immune lymphocytes important for host viral and tumor immunity. We investigated the requirement for ATP citrate lyase (ACLY) in NK cell function using an inducible genetic mouse model. ACLY regulates the citrate-malate shuttle, generating cytosolic acetyl-coenzyme A that is primarily used for acetylation or lipid synthesis. ACLY-deficient NK cells upon IL-15 activation exhibited significant defects in glycolysis, proliferation, cytokine production, and cytotoxicity, without decreased intracellular lipids. Notably, ACLY-deficiency specifically resulted in reduced NK cell responses to activating receptors associated with the adapter proteins DAP10 or DAP12. This is due to decreased DAP12 and increased DAP10 transcript and protein, coupled with epigenetic profiling that demonstrated altered histone acetylation of these genes in ACLY KO. Supplementation of ACLY-deficient NK cells with acetate was sufficient to overcome most functional defects, including restoring DAP10/12 expression and activating receptor function, emphasizing the importance of ACLY-generated cytosolic acetyl-coenzyme A for NK effector functions.

immunology↗

Big DNA as a tool to dissect an age-related macular degeneration haplotype

Age-related Macular Degeneration (AMD) is a leading cause of blindness in the developed world, especially in aging populations, and is therefore an important target for new therapeutic development. Recently, there have been several studies demonstrating strong associations between AMD and sites of heritable genetic variation at multiple loci, including a highly significant association at 10q26. The 10q26 risk region contains two genes, HTRA1 and ARMS2, both of which have been separately implicated as causative for the disease, as well as dozens of sites of non-coding variation. To date, no studies have successfully pinpointed which of these variant sites are functional in AMD, nor definitively identified which genes in the region are targets of such regulatory variation. In order to efficiently decipher which sites are functional in AMD phenotypes, we describe a general framework for combinatorial assembly of large synthetic haplotypes along with delivery to relevant disease cell types for downstream functional analysis. We demonstrate the successful and highly efficient assembly of a first-draft 119kb wild-type assemblon covering the HTRA1/ARMS2 risk region. We further propose the parallelized assembly of a library of combinatorial variant synthetic haplotypes covering the region, delivery and analysis of which will identify functional sites and their effects, leading to an improved understanding of AMD development. We anticipate that the methodology proposed here is highly generalizable towards the difficult problem of identifying truly functional variants from those discovered via GWAS or other genetic association studies.

genetics↗