Establishing comprehensive quaternary structural proteomes from genome sequence.

A critical body of knowledge has developed through advances in protein microscopy, protein-fold modeling, structural biology software, availability of sequenced bacterial genomes, large-scale mutation databases, and genome-scale models. Based on these recent advances, we develop a computational framework that; i) identifies the oligomeric structural proteome encoded by an organism's genome from available structural resources; ii) maps multi-strain alleleomic variation, resulting in the structural proteome for a species; and iii) calculates the 3D orientation of proteins across subcellular compartments with residue-level precision. Using the platform, we; iv) compute the quaternary E. coli K-12 MG1655 structural proteome; v) use a dataset of 12,000 mutations to build Random Forest classifiers that can predict the severity of mutations; and, in combination with a genome-scale model that computes proteome allocation, vi) obtain the spatial allocation of the E. coli proteome. Thus, in conjunction with relevant datasets and increasingly accurate computational models, we can now annotate quaternary structural proteomes, at genome-scale, to obtain a molecular-level understanding of whole-cell functions. Significance: Advancements in experimental and computational methods have revealed the shapes of multi-subunit proteins. The absence of a unified platform that maps actionable datatypes onto these increasingly accurate structures creates a barrier to structural analyses, especially at the genome-scale. Here, we describe QSPACE, a computational annotation platform that evaluates existing resources to identify the best-available structure for each protein in a user's query, maps the 3D location of actionable datatypes ( e.g. , active sites, published mutations) onto the selected structures, and uses third-party APIs to determine the subcellular compartment of all amino acids of a protein. As proof-of-concept, we deployed QSPACE to generate the quaternary structural proteome of E. coli MG1655 and demonstrate two use-cases involving large-scale mutant analysis and genome-scale modelling.

Establishing comprehensive quaternary structural proteomes from genome sequence.

A critical body of knowledge has developed through advances in protein microscopy, protein-fold modeling, structural biology software, availability of sequenced bacterial genomes, large-scale mutation databases, and genome-scale models. Based on these recent advances, we develop a computational platform that; i) computes the oligomeric structural proteome encoded by an organism's genome; ii) maps multi-strain alleleomic variation, resulting in the structural proteome for a species; and iii) calculates the 3D orientation of proteins across subcellular compartments with angstrom-level precision. Using the platform, we; iv) compute the full quaternary E. coli K-12 MG1655 structural proteome; v) deploy structure-guided analyses to identify consequential mutations; and, in combination with a genome-scale model that computes proteome allocation, vi) obtain a draft 3D visualization of the proteome in a functioning cell. Thus, in conjunction with relevant datasets and computational models, we can now resolve genome-scale structural proteomes to obtain an angstrom-level understanding of whole-cell functions.

Whole-genome sequences from wild-type and laboratory-evolved strains define the alleleome and establish its hallmarks.

The genomic diversity across strains of a species forms the genetic basis for differences in their behavior. A large-scale assessment of sequence variation has been made possible by the growing availability of strain-specific whole-genome sequences (WGS) and with the advent of large-scale databases of laboratory-acquired mutations. We define the Escherichia coli "alleleome" through a genome-scale assessment of amino acid (AA) sequence diversity in open reading frames across 2,661 WGS from wild-type strains. We observe a highly conserved alleleome enriched in mutations unlikely to affect protein function. In contrast, 33,000 mutations acquired in laboratory evolution experiments result in more severe AA substitutions that are rarely achieved by natural selection. Large-scale assessment of the alleleome establishes a method for the quantification of bacterial allelic diversity, reveals opportunities for synthetic biology to explore novel sequence space, and offers insights into the constraints governing evolution.