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Szeliova, D.

Publications and source records attributed to Szeliova, D..

7 recordsLinked to original sources

Growth costs of suboptimal protein allocation in nonlinear models of growing cells

Microbial growth depends on how cells allocate their limited proteome among competing functions. Although many proteins are expressed near levels that maximize growth, deviations from these optima are common and can impose substantial fitness costs. Modeling the growth costs of suboptimal protein allocation remains challenging because protein expression influences growth through multiple interacting mechanisms, including biosynthetic demands, enzyme kinetics, and limits on cellular density. Here we analyze these effects using growth balance analysis (GBA), a nonlinear framework that predicts steady-state growth and biomass composition of coarse-grained microbial models from basic cellular constraints. The optimal biomass compositions predicted for these models include not only protein concentrations but also the reactant concentrations required to saturate enzymes through nonlinear rate laws, thereby capturing more complex resource trade-offs than in conventional linear resource allocation models. Using these GBA models, we compute optimal biomass compositions with individual proteins fixed at suboptimal levels. The resulting relationships between suboptimal protein allocation and growth rate are consistent with qualitative experimental patterns in bacteria, with growth effects depending on protein function (including idle proteins), environmental conditions, toxic byproducts, and alternative reactions. These results indicate a practical and tractable approach for modeling growth costs arising from suboptimal protein allocation, and suggest a basis for predictive modeling in metabolic engineering and synthetic biology.

systems biology↗

The geometry of cooperation: decoding microbial interactions

Bridging the gap between mechanistic models of metabolism and ecological theory remains a key challenge in understanding interactions within microbial communities. We propose a geometric framework for analyzing community metabolism, based on constraint-based modeling. By extending pathway analysis methods from single organisms to multi-species systems, we define the community metabolic space as the set of all feasible fluxes between species and their environment, conditional on growth rate and medium composition. Embedded within a nonlinear geometry, this space forms a polytope whose vertices represent the minimal building blocks of community metabolism from which every feasible solution can be constructed. Strikingly, these elementary community flux modes invite direct ecological interpretation - as specialist, commensalist, or mutualist modes of cooperation. Furthermore, we find that mutualism is either isotypic (arising from a minimal mutualistic behavior) or anisotypic (emerging from a combination of reciprocal commensalists). This distinction demonstrates that bidirectional cross-feeding alone is insufficient to determine the ecological interaction type. Our framework also offers significant potential for applications. Because it does not rely on optimization, powerful unbiased tools from metabolic engineering, such as production envelopes and minimal cut sets, may be extended to microbial communities. Taken together, this perspective aims to unify ecological and metabolic viewpoints by linking interaction types to the geometric structure of the community metabolic space, thereby laying the foundation for a deeper understanding of community structure, function, and design.

systems biology↗

Elementary vectors reveal minimal interactions in microbial communities

Understanding microbial communities is essential for progress in ecology, biotechnology, and human health. In the last decade, constraint-based metabolic models of individual organisms have been combined to study microbial consortia. In this work, we present a geometric framework for characterizing all feasible microbial interactions. We project community models onto the key variables of interaction: exchange fluxes and community compositions. Based on this projection, we compute elementary composition/exchange fluxes (ECXs), extending the concept of minimal metabolic pathways from individual species to entire communities. Every feasible metabolic state of a community can be expressed as a combination of these elementary vectors. Notably, each ECX corresponds to a distinct ecological interaction type, such as specialization, commensalism, or mutualism. Finally, our geometric formulation enables the direct application of existing constraint-based methods, such as flux variability analysis and minimal cut sets, to microbial communities, providing a foundation for rational community design.

systems biology↗

Improving HEK293-based AAV-production using GSMMs, and a multi-omics approach

HEK293 cells are a versatile cell line extensively used in the production of recombinant proteins and viral vectors, notably Adeno-associated virus (AAV) [12]. Despite their high transfection efficiency and adaptability to various culture conditions, challenges remain in achieving sufficient yields of active viral particles. This study presents a comprehensive multi-omics analysis of two HEK293 strains under good manufacturing practice conditions, focusing on the metabolic and cellular responses during AAV production. The investigation included lipidomic, exometabolomic, and transcriptomic profiling across different conditions and time points. Genome-scale metabolic models (GSMMs) were reconstructed for these strains to elucidate metabolic shifts and identify potential bottlenecks in AAV production. Notably, the study revealed significant differences between a High-producing (HP) and a Low-producing (LP) HEK293 strains, highlighting pseudohypoxia in the LP strain. Key findings include the identification of hypoxia-inducible factor 1-alpha (HIF1alpha) as a critical regulator in the LP strain, linking pseudohypoxia to poor AAV productivity. Inhibition of HIF1alpha resulted in immediate cessation of cell growth and a 2-fold increase in viral capsid production, albeit with a decreased number of viral genomes, impacting the full-to-empty particle ratio. This suggests that while HIF1alpha inhibition enhances capsid assembly, it simultaneously hampers nucleotide synthesis via the pentose phosphate pathway (PPP), necessary for genome packaging.

bioengineering↗

Differential logistic PCA for large-scale comparison of GSMMs and pathway analysis

Genome-scale metabolic models (GSMMs) offer a holistic view of biochemical reaction networks, enabling in-depth analyses of metabolism across species and tissues in multiple conditions. However, comparing GSMMs against each other poses challenges as current dimensionality reduction algorithms or clustering methods lack mechanistic interpretability, and often rely on subjective assumptions. Here, we propose a new approach utilizing logisitic principal component analysis (LPCA) that efficiently clusters GSMMs while singling out mechanistic differences in terms of reactions and pathways that drive the categorization. We applied LPCA to multiple diverse datasets, including GSMMs of 222 Escherichia-strains, 343 budding yeasts (Saccharomycotina), 80 human tissues, and 2943 Firmicutes strains. Our findings demonstrate LPCAs effectiveness in preserving microbial phylogenetic relationships and discerning human tissue-specific metabolic profiles, exhibiting comparable performance to traditional methods like t-distributed stochastic neighborhood embedding (t-SNE) and Jaccard coefficients. Moreover, the subsystems and associated reactions identified by LPCA align with existing knowledge, underscoring its reliability in dissecting GSMMs and uncovering the underlying drivers of separation. Authors summaryGenome-scale metabolic models (GSMMs) are comprehensive representations of all the biochemical reactions that occur within an organism, enabling insights into cellular processes. Our study introduces logisitic principal component analysis (LPCA) to explore and compare these biochemical networks across different species and tissues only based on the presence or absence of reactions, summarized in a binary matrix. LPCA analyzes these binary matrices of specific biochemical reactions, identifying significant differences and similarities. We applied LPCA to a range of datasets, including bacterial strains, fungi, and human tissues. Our findings demonstrate LPCAs effectiveness in distinguishing microbial phylogenetic relationships and discerning tissue-specific profiles in humans. LPCA also offers precise information on the biochemical drivers of these differences, contributing to a deeper understanding of metabolic subsystems. This research showcases LPCA as a valuable method for examining the complex interplay of reactions within GSMMs, offering insights that could support further scientific investigation into metabolic processes.

systems biology↗

Does ribosome composition maximize growth rate? The role of RNA instability

Ribosomes are protein synthesis machines that are central to cellular self-fabrication, and the synthesis time of a ribosome places an upper bound on growth rate. While most cellular enzymes are proteins, ribosomes consist of 1/3 protein and 2/3 RNA (in E. coli). Recent research suggests that ribosome composition arises from a trade-off between two "autocatalytic loops", ribosomal protein and RNA polymerase synthesis, respectively. In this study, we develop a (coarse-grained) mechanistic model of a self-fabricating cell and validate it under various growth conditions. Using resource balance analysis (RBA), we examine how the maximum growth rate varies with ribosome composition, assuming that all kinetic parameters remain independent of ribosome composition. Our model highlights the importance of RNA instability. If we neglect it, RNA synthesis is always "cheaper" than protein synthesis, leading to an RNA-only ribosome at maximum growth rate. However, when we account for RNA turnover, we find that a mixed ribosome composed of RNA and proteins maximizes growth rate. To account for RNA turnover, we explore two scenarios regarding the activity of RNases. In (a) degradation is proportional to RNA content, whereas in (b) ribosomal proteins cooperatively mitigate RNA instability by protecting it from misfolding and subsequent degradation. In both cases, an increase in ribosomal protein content raises protein synthesis costs while reducing RNA turnover costs. This leads to a mixed ribosome composed of RNA and proteins. However, only in scenario (b), where we consider the cooperative protection of ribosomal RNA by proteins, our model predictions are in qualitative agreement with experimental data under different growth conditions. Our research offers new mechanistic insights into ribosome biogenesis and evolution. Furthermore, it paves the way for understanding the protein-rich ribosome composition found in archaea and mitochondria.

systems biology↗

Elementary vectors and autocatalytic sets for computational models of cellular growth

Traditional (genome-scale) metabolic models of cellular growth involve an approximate biomass "reaction", which specifies biomass composition in terms of precursor metabolites (such as amino acids and nucleotides). On the one hand, biomass composition is often not known exactly and may vary drastically between conditions and strains. On the other hand, the predictions of computational models crucially depend on biomass. Also elementary flux modes (EFMs), which generate the flux cone, depend on the biomass reaction. To better understand cellular phenotypes across growth conditions, we introduce and analyze new classes of elementary vectors for comprehensive (next-generation) metabolic models, involving explicit synthesis reactions for all macromolecules. Elementary growth modes (EGMs) are given by stoichiometry and generate the growth cone. Unlike EFMs, they are not support-minimal, in general, but cannot be decomposed "without cancellations". In models with additional (capacity) constraints, elementary growth vectors (EGVs) generate a growth polyhedron and depend also on growth rate. However, EGMs/EGVs do not depend on the biomass composition. In fact, they cover all possible biomass compositions and can be seen as unbiased versions of elementary flux modes/vectors (EFMs/EFVs) used in traditional models. To relate the new concepts to other branches of theory, we consider autocatalytic sets of reactions. Further, we illustrate our results in a small model of a self-fabricating cell, involving glucose and ammonium uptake, amino acid and lipid synthesis, and the expression of all enzymes and the ribosome itself. In particular, we study the variation of biomass composition as a function of growth rate. In agreement with experimental data, low nitrogen uptake correlates with high carbon (lipid) storage. Author summaryNext-generation, genome-scale metabolic models allow to study the reallocation of cellular resources upon changing environmental conditions, by not only modeling flux distributions, but also expression profiles of the catalyzing proteome. In particular, they do no longer assume a fixed biomass composition. Methods to identify optimal solutions in such comprehensive models exist, however, an unbiased understanding of all feasible allocations is missing so far. Here we develop new concepts, called elementary growth modes and vectors, that provide a generalized definition of minimal pathways, thereby extending classical elementary flux modes (used in traditional models with a fixed biomass composition). The new concepts provide an understanding of all possible flux distributions and of all possible biomass compositions. In other words, elementary growth modes and vectors are the unique functional units in any comprehensive model of cellular growth. As an example, we show that lipid accumulation upon nitrogen starvation is a consequence of resource allocation and does not require active regulation. Our work puts current approaches on a theoretical basis and allows to seamlessly transfer existing workflows (e.g. for the design of cell factories) to next-generation metabolic models.

systems biology↗