Large tree-of-life (ToL) scale databases of microbial genomes are powerful resources for exploring genome structure and function from a phylogenomic context. Despite the growing availability of genomic data, large-scale genome annotation is still a challenge, with a considerable fraction of genes remaining as unannotated. Here, to expand the capabilities of database-wide gene function prediction, we used our AnnoTree platform as a corpus for training a Word2vec-based genomic language model (gLM). Machine-learning of genomic grammar patterns across the AnnoTree database revealed functional associations between genes and enabled the inference of function for hypothetical proteins and domains, as we demonstrated by predicting novel type VI secretion proteins. Finally, we implemented a web-server to allow users to interact with the AnnoTree Word2vec model, thus facilitating gene function prediction. Ultimately, our work highlighted the GTDB/AnnoTree database as a powerful training database for gLMs focused on prediction and discovery of microbial gene functions.

The analysis and comparison of gene neighborhoods is a powerful approach for exploring microbial genome structure, function, and evolution. Although numerous tools exist for genome visualization and comparison, genome exploration across large genomic databases or user-generated datasets remains a challenge. Here, we introduce AnnoView, a web server designed for interactive exploration of gene neighborhoods across the bacterial and archaeal tree of life. Our server offers users the ability to identify, compare, and visualize gene neighborhoods of interest from 30 238 bacterial genomes and 1672 archaeal genomes, through integration with the comprehensive Genome Taxonomy Database and AnnoTree databases. Identified gene neighborhoods can be visualized using pre-computed functional annotations from different sources such as KEGG, Pfam and TIGRFAM, or clustered based on similarity. Alternatively, users can upload and explore their own custom genomic datasets in GBK, GFF or CSV format, or use AnnoView as a genome browser for relatively small genomes (e.g. viruses and plasmids). Ultimately, we anticipate that AnnoView will catalyze biological discovery by enabling user-friendly search, comparison, and visualization of genomic data. AnnoView is available at http://annoview.uwaterloo.ca.

MOTIVATION: Analyses in comparative genomics often require non-redundant genome datasets. Eliminating redundancy is not as simple as keeping one strain for each named species because genomes might be redundant at a higher taxonomic level than that of species for some analyses; some strains with different species names can be as similar as most strains sharing a species name, whereas some strains sharing a species name can be so different that they should be put into different groups; and some genomes lack a species name. RESULTS: We have implemented a method and Web server that clusters a genome dataset into groups of redundant genomes at different thresholds based on a few phylogenomic distance measures. AVAILABILITY: The Web interface, similarity and distance data and R-scripts can be accessed at http://microbiome.wlu.ca/research/redundancy/.

MOTIVATION: The analyses of the increasing number of genome sequences requires shortcuts for the detection of orthologs, such as Reciprocal Best Hits (RBH), where orthologs are assumed if two genes each in a different genome find each other as the best hit in the other genome. Two BLAST options seem to affect alignment scores the most, and thus the choice of a best hit: the filtering of low information sequence segments and the algorithm used to produce the final alignment. Thus, we decided to test whether such options would help better detect orthologs. RESULTS: Using Escherichia coli K12 as an example, we compared the number and quality of orthologs detected as RBH. We tested four different conditions derived from two options: filtering of low-information segments, hard (default) versus soft; and alignment algorithm, default (based on matching words) versus Smith-Waterman. All options resulted in significant differences in the number of orthologs detected, with the highest numbers obtained with the combination of soft filtering with Smith-Waterman alignments. We compared these results with those of Reciprocal Shortest Distances (RSD), supposed to be superior to RBH because it uses an evolutionary measure of distance, rather than BLAST statistics, to rank homologs and thus detect orthologs. RSD barely increased the number of orthologs detected over those found with RBH. Error estimates, based on analyses of conservation of gene order, found small differences in the quality of orthologs detected using RBH. However, RSD showed the highest error rates. Thus, RSD have no advantages over RBH. AVAILABILITY: Orthologs detected as Reciprocal Best Hits using soft masking and Smith-Waterman alignments can be downloaded from http://popolvuh.wlu.ca/Orthologs.

Abstract Motivation: The prediction of the transcription unit organization of genomes is an important clue in the inference of functional relationships of genes, the interpretation and evaluation of transcriptome experiments, and the overall inference of the regulatory networks governing the expression of genes in response to the environment. Though several methods have been devised to predict operons, most need a high characterization of the genome analysed. Log-likelihoods derived from inter-genic distance distributions work surprisingly well to predict operons in Escherichia coli and are available for any genome as soon as the gene sets are predicted. Results: Here we provide evidence that the very same method is applicable to any prokaryotic genome. First, the method has the same efficiency when evaluated using a collection of experimentally known operons of Bacillus subtilis. Second, operons among most if not all prokaryotes seem to have the same tendencies to keep short distances between their genes, the most frequent distances being the overlaps of four and one base pairs. The universality of this structural feature allows us to predict the organization of transcription units in all prokaryotes. Third, predicted operons contain a higher proportion of genes with related phylogenetic profiles and conservation of adjacency than predicted borders of transcription units. Supplementary information: Additional materials and graphs, are available at: http://www.cifn.unam.mx/moreno/pub/TUpredictions/ Contact: [email protected] Keywords: functional genomics; comparative genomics; operon prediction; operon structure. *To whom correspondence should be addressed.