Biology subjects

Andre, K.

Publications and source records attributed to Andre, K..

3 recordsLinked to original sources

bioRxiv2024

Halofilins as Emerging Bactofilin Families of Archaeal Cell Shape Plasticity Orchestrators

Bactofilins are rigid, non-polar bacterial cytoskeletal filaments that link cellular processes to specific curvatures of the cytoplasmic membrane. Although homologs of bactofilins have been identified in archaea and eukaryotes, functional studies have remained confined to bacterial systems. Here, we characterize representatives of two new families of archaeal bactofilins from the pleomorphic archaeon Haloferax volcanii, halofilin A (HalA) and halofilin B (HalB). HalA and HalB polymerize in vitro, assembling into straight bundles. HalA polymers are highly dynamic and accumulate at positive membrane curvatures in vivo, whereas HalB forms more static foci that localize in areas of local negative curvatures on the outer cell surface. Gene deletions and live-cell imaging show that halofilins are critical in maintaining morphological integrity during shape transition from disk (sessile) to rod (motile). Morphological defects in {Delta}halA result in accumulation of highly positive curvatures in rods but not in disks. Conversely, disk-shaped cells are exclusively affected by halB deletion, resulting in flatter cells. Furthermore, while {Delta}halA and {Delta}halB cells imprecisely determine the future division plane, defects arise predominantly during the disk-to-rod shape remodeling. In fact, the deletion of halA in the haloarchaeon Halobacterium salinarum, whose cells are consistently rod-shaped, impacted morphogenesis but not cell division. Increased levels of halofilins enforced drastic deformations in cells devoid of S-layer, suggesting that HalB polymers are more stable at defective S-layer lattice regions. Our results set halofilins apart from their bacterial correlate, where they provide mechanical scaffolding instead of directing envelope synthesis.

microbiology↗

bioRxiv2023

A Genome-Wide Comprehensive Analysis of Nucleosome Positioning in Yeast

In eukaryotic cells, the one-dimensional DNA molecules need to be tightly packaged into the spatially constraining nucleus. Folding is achieved on its lowest level by wrapping the DNA around nucleosomes. Their positioning regulates other nuclear processes, such as transcription and DNA repair. Despite strong efforts to study nucleosome phasing using Next Generation Sequencing (NGS) data, the mechanism of their collective arrangement along the gene body remains poorly understood. Here, we assess the nucleosome profiles of protein-coding genes in Saccharomyces cerevisiae using functional Principal Component Analysis. By decomposing the NGS signals into their main descriptive functions, we compared wild type and chromatin remodeler-deficient strains, keeping position-specific details preserved. A correlation analysis with other genomic properties, such as gene size and length of the upstream Nucleosome Depleted Region (NDR), identified key factors that influence nucleosome phasing. We reveal that the RSC chromatin remodeler--which is responsible for NDR maintenance--is indispensable for decoupling nucleosome arrangement within the gene from phasing outside, which interfere in rsc8-depleted conditions. Moreover, positioning in chd1{Delta} strains displayed a clear correlation with RNA polymerase II presence, whereas wild type cells did not indicate a noticeable interdependence. We propose that RSC is pivotal for global nucleosome organisation, whilst Chd1 plays a key role for maintaining local arrangement.

bioinformatics↗

bioRxiv2023

A sweet new set of inducible and constitutive promoters for haloarchaea

Inducible promoters are one of cellular and molecular biologys most important technical tools. The ability to deplete, replete, and overexpress genes on demand is the foundation of most functional studies. Here, we developed and characterized a new xylose-responsive promoter (Pxyl), the second inducible promoter system for the model haloarcheon Haloferax volcanii. Generating RNA-seq datasets from cultures in the presence of four historically used inducers (arabinose, xylose, maltose, and IPTG), we mapped upregulated genomic regions primarily repressed in the absence of the above inducers. We found a highly upregulated promoter that controls the expression of the xacEA (HVO_B0027-28) operon in the pHV3 chromosome. To characterize this promoter region, we cloned msfGFP (monomeric superfold green fluorescent protein) under the control of two different 5 UTR fragments into a modified pTA962 vector: the first 250 bp (P250) and the whole 750 bp intergenic region (P750). The P250 region expressed msfGFP constitutively, and its expression did not respond to the presence or absence of xylose. However, the P750 promoter showed not only to be repressed in the absence of xylose but also expressed higher levels of msfGFP than the previously described inducible promoter PtnaA in the presence of the inducer. Finally, we validated the inducible Pxyl promoter by reproducing morphological phenotypes already described in the literature. By overexpressing the tubulin-like FtsZ1 and FtsZ2, we observed similar but slightly more pronounced morphological defects than the tryptophan-inducible promoter PtnaA. FtsZ1 overexpression created larger, deformed cells, whereas cells overexpressing FtsZ2 were smaller but mostly retained their shape. In summary, this work contributes a new xylose-inducible promoter, Pxyl, that can be used simultaneously with the well-established PtnaA in functional studies in H. volcanii.

microbiology↗