Biology subjects

Steube, N.

Publications and source records attributed to Steube, N..

3 recordsLinked to original sources

bioRxiv2025

Spatiotemporal control of cortical centrin patterning by regionalized Sfi1 family scaffolding proteins in Stentor coeruleus

Summary/AbstractPatterning is fundamental to the development and maintenance of organisms, ensuring functional and structural organization. While patterning is well studied at the level of multicellular organisms, even single cells need to undergo morphogenesis and form spatial patterns. Stentor coeruleus is a large ciliate that has been a classical system for studying patterning and morphogenesis due to its distinctive shape and organization of easily visible cortical structures which show a clear localization along the anterior-posterior body axis. The cortex of Stentor is made of two cytoskeletal layers, one composed of microtubules, and one composed of a network of centrin-family EF hand proteins, which form a branched network in the anterior half of the cell and long thick bundles known as myonemes in the posterior half. Sfi1 family proteins scaffold the assembly of centrin filaments throughout the eukaryotes, and function as scaffolding proteins in myonemal contractile systems found in some ciliates. A set of Sfi1 proteins upregulated during regeneration in Stentor were found to have a role in maintaining anterior/posterior differences in centrin patterning. Loss of this distinct anterior-posterior patterning leads to defects in regeneration and contraction. Using RNAi-mediated knockdown of Sfi1 genes, we have found that cells fail to regenerate their oral apparatus, with early expressed Sfi1 genes more critical for oral primordium (assembly of organized basal bodies that become the new oral apparatus) formation than those expressed later. Additionally, different Sfi1 proteins appear to be recruited sequentially to the growing primordium in an order that matches the time at which they are required for progression of regeneration. Knockdown of Sfi1 isoforms that results in reduced myoneme cables impairs contraction such that cells fail to contract in response to a stimulus or do not fully contract. These findings suggest a model in which regionalized differences in patterning of cytoskeletal assemblies can be modulated by regionalized localization of scaffolding proteins.

cell biology↗

bioRxiv2022

ATPase activity of B. subtilis RecA affects the dynamic formation of RecA filaments at DNA double strand breaks

RecA plays a central role in DNA repair and is a main actor involved in homologous recombination (HR). In vivo, RecA forms filamentous structures termed "threads", which are essential for HR, but whose nature is still ill defined. We show that RecA from Bacillus subtilis having lower ATP binding activity can still form nucleoprotein filaments in vitro, and still retains most of wild type RecA activity in vivo. Contrarily, loss of ATPase activity strongly reduces formation of nucleoprotein filaments in vitro, and effectivity to repair double strand breaks (DSBs) in vivo. While lowered ATP-binding activity only moderately affected RecA dynamics, loss of ATPase activity lead to a large reduction of the formation of threads, as well as of their dynamic changes observed in a seconds-scale. Single molecule tracking of RecA revealed incorporation of freely diffusing and non-specifically DNA-bound molecules into filaments upon induction of a single DSB. This change of dynamics was highly perturbed in the absence of ATPase activity, revealing that filamentous forms of RecA as well as their dynamics depend on ATPase activity. Our data suggest that RecA/ssDNA filaments change in subcellular localization and length involving ATP-driven homology search.

microbiology↗

bioRxiv2021

Green light perception paved the way for the diversification of GAF domain photoreceptors

Photoreceptors are proteins that sense incident light and then trigger downstream signaling events. Phytochromes are linear tetrapyrrole-binding photoreceptors present in plants, algae, fungi, and various bacteria. Most phytochromes respond to red and far-red light signals. Among the phytochrome superfamily, cyanobacteria-specific cyanobacteriochromes show much more diverse optical properties covering the entire visible region. Both phytochromes and cyanobacteriochromes share the GAF domain scaffold to cradle the chromophore as the light-sensing region. It is unknown what physiological demands drove the evolution of cyanobacteriochromes in cyanobacteria. Here we utilize ancestral sequence reconstruction and report that the resurrected ancestral cyanobacteriochrome proteins reversibly respond to green and red light signals. pH titration analyses indicate that the deprotonation of the bound phycocyanobilin chromophore enables the photoreceptor to perceive green light. The ancestral cyanobacteriochromes show modest thermal reversion to the green light-absorbing form, suggesting that they evolved to sense green-rich irradiance rather than red light, which is preferentially utilized for photosynthesis. In contrast to plants and green algae, many cyanobacteria can utilize green light for photosynthesis with their special light-harvesting complexes, phycobilisomes. The evolution of green/red sensing cyanobacteriochromes may therefore have allowed ancient cyanobacteria to acclimate to different light environments by rearranging the absorption capacity of the cyanobacterial antenna complex by chromatic acclimation. Significance StatementLight serves as a crucial environmental stimulus affecting the physiology of organisms across all kingdoms of life. Photoreceptors serve as important players of light responses, absorbing light and actuating biological processes. Among a plethora of photoreceptors, cyanobacteriochromes arguably have the wealthiest palette of color sensing, largely contributing to the success of cyanobacteria in various illuminated habitats. Our ancestral sequence reconstruction and the analysis of the resurrected ancestral proteins suggest that the very first cyanobacteriochrome most probably responded to the incident green-to-red light ratio, in contrast to modern red/far-red absorbing plant phytochromes. The deprotonation of the light-absorbing pigment for green light-sensing was a crucial molecular event for the invention of the new class of photoreceptors with their huge color tuning capacity.

biochemistry↗