Biology subjects

Vlahovicek, K.

Publications and source records attributed to Vlahovicek, K..

8 recordsLinked to original sources

bioRxiv2026

Hidden Contaminants in Sponge Genomes: Large-Scale Decontamination of 30 Public Assemblies

Sponges (phylum Porifera) are early-diverging metazoans that play central ecological roles and serve as models for understanding animal evolution. However, their associations with diverse microbial communities increase the risk of contamination in publicly available datasets, potentially compromising downstream biological inference. Despite growing genomic resources, systematic assessments of contamination in sponge genome assemblies have been lacking. Here, we present a comprehensive contamination analysis of 30 publicly available sponge genome assemblies and introduce a reproducible and easily adoptable decontamination pipeline tailored to non-model organisms. Using this framework, we provide decontaminated versions of the analysed assemblies. The pipeline integrates three complementary lines of evidence: compositional outlier detection based on k-mer profiles and GC content, protein-level taxonomic classification using DIAMOND, and nucleotide-level classification with Kraken2. Scaffolds are designated as contaminants when supported by at least two independent signals. Pipeline performance was validated using a realistic spike-in dataset composed of bona fide sponge sequences and representative contaminant genomes. The decontamination pipeline achieved 96.8% accuracy, 99.6% precision, and 90.8% recall, maintaining consistently strong recall across the vast majority of analyzed taxa. In addition, taxonomic assignments were accurately resolved to the genus level for 96.3% of identified contaminants. Application to public assemblies revealed variable contamination. On average, 14.5% of scaffolds per assembly were classified as contaminants, although they represented a low fraction of the total genome length, indicating that contamination is concentrated in relatively short scaffolds. Detected contaminants were dominated by bacterial phyla commonly associated with sponge microbiomes, including Pseudomonadota, Chloroflexota, and Poribacteria, with additional archaeal, protozoan, algal, and fungal sequences. Importantly, the number of complete BUSCO orthologs remained virtually unchanged following contamination removal, indicating minimal loss of genuine host scaffolds. Taken together, our study provides 30 curated sponge genome assemblies and a consensus-based decontamination framework tailored to non-model organisms, improving the reliability of genomic resources for evolutionary, ecological, and functional analyses.

genomics↗

bioRxiv2025

Comprehensive Transcriptome Quality Assessment Using CATS: Reference-free and Reference-based Approaches

Accurate assessment of transcriptome assembly quality is critical to ensure the reliability of subsequent transcriptomic analyses. We present CATS (Comprehensive Assessment of Transcript Sequences), a tool offering both reference-free (CATS-rf) and reference-based (CATS-rb) transcriptome quality evaluation pipelines. CATS-rf maps RNA-seq reads back to the assembled transcripts and computes four interpretable scoring components that capture common assembly errors. CATS-rb assesses transcriptome completeness via alignment to a reference genome, supporting both annotation-free and annotation-based scoring. We benchmarked CATS on 672 transcriptomes from simulated and public RNA-seq data. CATS-rf outperformed existing tools in both transcript-level accuracy assessment and demonstrated high sensitivity to diverse assembly error types. CATS-rb produced robust transcriptome completeness estimates even without external annotation, with its scoring metrics strongly reflecting assembly quality. These results highlight CATS as an accurate, interpretable, and broadly applicable framework for evaluating transcriptome assemblies.

bioinformatics↗

bioRxiv2024

Somatic embryogenesis of grapevine (Vitis vinifera) expresses a transcriptomic hourglass

At the molecular level, multicellular eukaryotic lineages and bacterial biofilms show predictable evolutionary footprints in their development. For instance, the zygotic embryogenesis of Arabidopsis, which is initiated by gamete fusion, shows hourglass-shaped ontogeny-phylogeny correlations at the transcriptome level. However, many plants are capable of yielding a fully viable next generation by somatic embryogenesis -- a comparable developmental process that usually starts by the embryogenic induction of a diploid somatic cell. This leads to the question: is the hourglass-shaped ontogeny-phylogeny correlation preserved in somatic embryogenesis? To explore the correspondence between ontogeny and phylogeny in this alternative developmental route in plants, we developed a new and highly efficient model of somatic embryogenesis in grapevine (Vitis vinifera) and sequenced its developmental transcriptomes. By combining the evolutionary properties of grapevine genes with their expression values, which were recovered from early induction until the formation of juvenile plants, we found a strongly supported hourglass-shaped developmental trajectory. However, in contrast to zygotic embryogenesis in Arabidopsis where the torpedo stage was evolutionary the most inert, we found that in the somatic embryogenesis of grapevine the heart stage expressed evolutionary the oldest and the most conserved transcriptome. This is a surprising finding because it suggests a better evolutionary system-level analogy between animal development and plant somatic embryogenesis than zygotic embryogenesis. We conclude that macroevolutionary logic is deeply hardwired in plant ontogeny and that somatic embryogenesis is likely a primordial embryogenic program in plants.

evolutionary biology↗

bioRxiv2022

Genomic profiling of HIV-1 integration in microglia links viral insertions to TAD organization

HIV-1 persists in anatomically distinct cellular and tissue reservoirs as a stably integrated provirus that is a major barrier to HIV-1 cure. Proviral insertions are largely characterized in blood cells, while HIV-1 integration patterns remain unexplored in microglia, the major brain reservoir. Here, we employ genomics approaches to obtain the first HIV-1 integration site (IS) profiling in microglia and perform in-depth analysis of transcriptome, specific histone signatures and chromatin accessibility on different genomic scales. We show that HIV-1 follows genic insertion patterns into introns of actively transcribed genes, characteristic of blood reservoirs. HIV-1 insertional hotspot analysis by non-negative matrix factorization (NMF)-based approach clusters IS signatures with genic- and super-enhancers. Chromatin accessibility transcription factor (TF) footprints reveal that increased CTCF binding marks latently infected microglia compared to productively infected one. We identify CTCF-enriched topologically associated domain (TAD) borders with signatures of active chromatin as a neighborhood for HIV-1 integration in microglia and CD4+ T cells. Our findings further strengthen the notion that HIV-1 follows the patterns of host cell genome organization to integrate and to establish the silent proviral state and reveal that these principles are largely conserved in different anatomical latent reservoirs.

genomics↗

bioRxiv2020

Embryo-like features in developing Bacillus subtilis biofilms

Correspondence between evolution and development has been discussed for more than two centuries1. Recent work reveals that phylogeny-ontogeny correlations are indeed present in developmental transcriptomes of eukaryotic clades with complex multicellularity2-10. Nevertheless, it has been largely ignored that the pervasive presence of phylogeny-ontogeny correlations is a hallmark of development in eukaryotes6, 10-12. This perspective opens a possibility to look for similar parallelisms in biological settings where developmental logic and multicellular complexity are more obscure13-16. For instance, it has been increasingly recognized that multicellular behaviour underlies biofilm formation in bacteria13, 14, 17-19. However, it remains unclear whether bacterial biofilm growth shares some basic principles with development in complex eukaryotes14-16, 18, 20. Here we show that the ontogeny of growing Bacillus subtilis biofilms recapitulates phylogeny at the expression level. Using time-resolved transcriptome and proteome profiles, we found that biofilm ontogeny correlates with the evolutionary measures, in a way that evolutionary younger and more diverged genes were increasingly expressed towards later timepoints of biofilm growth. Molecular and morphological signatures also revealed that biofilm growth is highly regulated and organized into discrete ontogenetic stages, analogous to those of eukaryotic embryos11, 21. Together, this suggests that biofilm formation in Bacillus is a bona fide developmental process comparable to organismal development in animals, plants and fungi. Given that most cells on Earth reside in the form of biofilms22 and that biofilms represent the oldest known fossils23, we anticipate that the widely-adopted vision of the first life as a single-cell and free-living organism needs rethinking.

evolutionary biology↗

bioRxiv2019

Tumor mutational landscape is a record of the pre-malignant state

Chromatin structure has a major influence on the cell-specific density of somatic mutations along the cancer genome. Here, we present a pan-cancer study in which we searched for the putative cancer cell-of-origin of 2,550 whole genomes, representing 32 cancer types by matching their mutational landscape to the regional patterns of chromatin modifications ascertained in 104 normal tissue types. We found that, in almost all cancer types, the cell-of-origin can be predicted solely from their DNA sequences. Our analysis validated the hypothesis that high-grade serous ovarian cancer originates in the fallopian tube and identified distinct origins of breast cancer subtypes. We also demonstrated that the technique is equally capable of identifying the cell-of-origin for a series of 2,044 metastatic samples from 22 of the tumor types available as primaries. Moreover, cancer drivers, whether inherited or acquired, reside in active chromatin regions in the respective cell-of-origin. Taken together, our findings highlight that many somatic mutations accumulate while the chromatin structure of the cell-of-origin is maintained and that this historical record, captured in the DNA, can be used to identify the often elusive cancer cell-of-origin.

genomics↗

bioRxiv2018

Role of Cnot6l in maternal mRNA turnover

Removal of poly(A) tail is an important mechanism controlling eukaryotic mRNA turnover. The major eukaryotic deadenylase complex CCR4-NOT contains two deadenylase components, CCR4 and CAF1 for which mammalian CCR4 is encoded by Cnot6 or Cnot6l paralogs. We show that Cnot6l apparently supplies the majority of CCR4 in the maternal CCR4-NOT complex in mouse, hamster, and bovine oocytes. Deletion of Cnot6l yielded viable mice but Cnot6l-/- females exhibited ~40% smaller litter size. The main onset of the phenotype was post-zygotic: fertilized Cnot6l-/- eggs developed slower and arrested more frequently than Cnot6l+/- eggs suggesting that maternal CNOT6L is necessary for accurate oocyte-to-embryo transition (OET). Transcriptome analysis revealed major transcriptome changes in Cnot6l-/- ovulated eggs and 1-cell zygotes. In contrast, minimal transcriptome changes in preovulatory Cnot6l-/- oocytes were consistent with reported Cnot6l mRNA dormancy. A minimal overlap between transcripts sensitive to decapping inhibition and Cnot6l loss suggests that decapping and CNOT6L-mediated deadenylation selectively target distinct subsets of mRNAs during OET in mouse.

molecular biology↗

bioRxiv2018

Spatially clustered loci with multiple enhancers are frequent targets of HIV-1

HIV-1 recurrently targets active genes that are positioned in the outer shell of the nucleus and integrates in the proximity of the nuclear pore compartment. However, the genomic features of these genes and the relevance of their transcriptional activity for HIV-1 integration have so far remained unclear. Here we show that recurrently targeted genes are delineated with super-enhancer genomic elements and that they cluster in specific spatial compartments of the T cell nucleus. We further show that these gene clusters acquire their location at the nuclear periphery during the activation of T cells. The clustering of these genes along with their transcriptional activity are the major determinants of HIV-1 integration in T cells. Our results show for the first time the relevance of the spatial compartmentalization of the genome for HIV-1 integration, thus further strengthening the role of nuclear architecture in viral infection.

genomics↗