SARS-CoV-2 receptor-binding domain deep mutational AlphaFold2 structures.

Leveraging recent advances in computational modeling of proteins with AlphaFold2 (AF2) we provide a complete curated data set of all single mutations from each of the 7 main SARS-CoV-2 lineages spike protein receptor binding domain (RBD) resulting in 3819X7 = 26733 PDB structures. We visualize the generated structures and show that AF2 pLDDT values are correlated with state-of-the-art disorder approximations, implying some internal protein dynamics are also captured by the model. Joint increasing mutational coverage of both structural and phenotype data coupled with advances in machine learning can be leveraged to accelerate virology research, specifically future variant prediction. We hope this data release can offer assistance into further understanding of the local and global mutational landscape of SARS-CoV-2 as well as provide insight into the biological understanding that 3D structure acts as a bridge between protein genotype and phenotype.

Identification of mutations in SARS-CoV-2 PCR primer regions.

Due to the constantly increasing number of mutations in the SARS-CoV-2 genome, concerns have emerged over the possibility of decreased diagnostic accuracy of reverse transcription-polymerase chain reaction (RT-PCR), the gold standard diagnostic test for SARS-CoV-2. We propose an analysis pipeline to discover genomic variations overlapping the target regions of commonly used PCR primer sets. We provide the list of these mutations in a publicly available format based on a dataset of more than 1.2 million SARS-CoV-2 samples. Our approach distinguishes among mutations possibly having a damaging impact on PCR efficiency and ones anticipated to be neutral in this sense. Samples are categorized as "prone to misclassification" vs. "likely to be correctly detected" by a given PCR primer set based on the estimated effect of mutations present. Samples susceptible to misclassification are generally present at a daily rate of 2% or lower, although particular primer sets seem to have compromised performance when detecting Omicron samples. As different variant strains may temporarily gain dominance in the worldwide SARS-CoV-2 viral population, the efficiency of a particular PCR primer set may change over time, therefore constant monitoring of variations in primer target regions is highly recommended.

Origin of Increased Solvent Accessibility of Peptide Bonds in Mutual Synergetic Folding Proteins.

Mutual Synergetic Folding (MSF) proteins belong to a recently discovered class of proteins. These proteins are disordered in their monomeric but ordered in their oligomeric forms. Their amino acid composition is more similar to globular proteins than to disordered ones. Our preceding work shed light on important structural aspects of the structural organization of these proteins, but the background of this behavior is still unknown. We suggest that solvent accessibility is an important factor, especially solvent accessibility of the peptide bonds can be accounted for this phenomenon. The side chains of the amino acids which form a peptide bond have a high local contribution to the shielding of the peptide bond from the solvent. During the oligomerization step, other non-local residues contribute to the shielding. We investigated these local and non-local effects of shielding based on Shannon information entropy calculations. We found that MSF and globular homodimeric proteins have different local contributions resulting from different amino acid pair frequencies. Their non-local distribution is also different because of distinctive inter-subunit contacts.

Analysis of Heterodimeric "Mutual Synergistic Folding"-Complexes.

Several intrinsically disordered proteins (IDPs) are capable to adopt stable structures without interacting with a folded partner. When the folding of all interacting partners happens at the same time, coupled with the interaction in a synergistic manner, the process is called Mutual Synergistic Folding (MSF). These complexes represent a discrete subset of IDPs. Recently, we collected information on their complexes and created the MFIB (Mutual Folding Induced by Binding) database. In a previous study, we compared homodimeric MSF complexes with homodimeric and monomeric globular proteins with similar amino acid sequence lengths. We concluded that MSF homodimers, compared to globular homodimeric proteins, have a greater solvent accessible main-chain surface area on the contact surface of the subunits, which becomes buried during dimerization. The main driving force of the folding is the mutual shielding of the water-accessible backbones, but the formation of further intermolecular interactions can also be relevant. In this paper, we will report analyses of heterodimeric MSF complexes. Our results indicate that the amino acid composition of the heterodimeric MSF monomer subunits slightly diverges from globular monomer proteins, while after dimerization, the amino acid composition of the overall MSF complexes becomes more similar to overall amino acid compositions of globular complexes. We found that inter-subunit interactions are strengthened, and additionally to the shielding of the solvent accessible backbone, other factors might play an important role in the stabilization of the heterodimeric structures, likewise energy gain resulting from the interaction of the two subunits with different amino acid compositions. We suggest that the shielding of the ß-sheet backbones and the formation of a buried structural core along with the general strengthening of inter-subunit interactions together could be the driving forces of MSF protein structural ordering upon dimerization.

Siculibacillus lacustris gen. nov., sp. nov., a new rosette-forming bacterium isolated from a freshwater crater lake (Lake St. Ana, Romania).

A new aerobic alphaproteobacterium, strain SA-279T, was isolated from a water sample of a crater lake. The 16S rRNA gene sequence analysis revealed that strain SA-279T formed a distinct lineage within the family Ancalomicrobiaceae and shared the highest pairwise similarity values with Pinisolibacterravus E9T (96.4 %) and Ancalomicrobiumadetum NBRC 102456T (94.2 %). Cells of strain SA-279T were rod-shaped, motile, oxidase and catalase positive, and capable of forming rosettes. Its predominant fatty acids were C18 : 1ω7c (69.0 %) and C16 : 1ω7c (22.7 %), the major respiratory quinone was Q-10, and the main polar lipids were phosphatidylethanolamine, phosphatidylmonomethylethanolamine, phosphatidylcholine, phosphatidylglycerol, an unidentified aminophospholipid and an unidentified lipid. The G+C content of the genomic DNA of strain SA-279T was 69.2 mol%. On the basis of the phenotypic, chemotaxonomic and molecular data, strain SA-279T is considered to represent a new genus and species within the family Ancalomicrobiaceae, for which the name Siculibacillus lacustris gen. nov., sp. nov. is proposed. The type strain is SA-279T (=DSM 29840T=JCM 31761T).

Physical Background of the Disordered Nature of "Mutual Synergetic Folding" Proteins.

Intrinsically disordered proteins (IDPs) lack a well-defined 3D structure. Their disordered nature enables them to interact with several other proteins and to fulfil their vital biological roles, in most cases after coupled folding and binding. In this paper, we analyze IDPs involved in a new mechanism, mutual synergistic folding (MSF). These proteins define a new subset of IDPs. Recently we collected information on these complexes and created the Mutual Folding Induced by Binding (MFIB) database. These protein complexes exhibit considerable structural variation, and almost half of them are homodimers, but there is a significant amount of heterodimers and various kinds of oligomers. In order to understand the basic background of the disordered character of the monomers found in MSF complexes, the simplest part of the MFIB database, the homodimers are analyzed here. We conclude that MFIB homodimeric proteins have a larger solvent-accessible main-chain surface area on the contact surface of the subunits, when compared to globular homodimeric proteins. The main driving force of the dimerization is the mutual shielding of the water-accessible backbones and the formation of extra intermolecular interactions.

A new Rhizobium species isolated from the water of a crater lake, description of Rhizobium aquaticum sp. nov.

A novel isolate, strain SA-276T, was isolated from the water of Lake St. Ana, a crater lake which is located in Romania. Phylogenetic analysis based on the 16S rRNA gene revealed that the new strain is a member of the family Rhizobiaceae, showing a high pairwise similarity value (97.65%) to Rhizobium tubonense CCBAU 85046T (= DSM 25379T), Rhizobium leguminosarum USDA 2370T (= LMG 14904T), Rhizobium anhuiense CCBAU 23252T and Rhizobium laguerreae FB206T. Cells of strain SA-276T were rod-shaped, motile, oxidase negative and weakly catalase positive. The predominant fatty acids were C18:1ω7c and cyclo C19:0ω8c, the major respiratory quinones were Q-10 and Q-9, and the main polar lipids were phosphatidylmonomethylethanolamine, phosphatidylglycerol and phosphatidylcholine. The G + C content of the genomic DNA of strain SA-276T was 60.8 mol%. The novel isolate can be distinguished from the closest related type strain R. tubonense DSM 25379T based on its broader substrate specificity and positive trypsin enzyme activity. On the basis of the phenotypic, chemotaxonomic and molecular data, strain SA-276T is considered to represent a new species, for which the name Rhizobium aquaticum sp. nov. is proposed. The type strain is SA-276T (= DSM 29780T = JCM 31760T).

Differences in planktonic microbial communities associated with three types of macrophyte stands in a shallow lake.

Little is known about how various substances from living and decomposing aquatic macrophytes affect the horizontal patterns of planktonic bacterial communities. Study sites were located within Lake Kolon, which is a freshwater marsh and can be characterised by open-water sites and small ponds with different macrovegetation (Phragmites australis, Nymphea alba and Utricularia vulgaris). Our aim was to reveal the impact of these macrophytes on the composition of the planktonic microbial communities using comparative analysis of environmental parameters, microscopy and pyrosequencing data. Bacterial 16S rRNA gene sequences were dominated by members of phyla Proteobacteria (36%-72%), Bacteroidetes (12%-33%) and Actinobacteria (5%-26%), but in the anoxic sample the ratio of Chlorobi (54%) was also remarkable. In the phytoplankton community, Cryptomonas sp., Dinobryon divergens, Euglena acus and chrysoflagellates had the highest proportion. Despite the similarities in most of the measured environmental parameters, the inner ponds had different bacterial and algal communities, suggesting that the presence and quality of macrophytes directly and indirectly controlled the composition of microbial plankton.

Quisquiliibacterium transsilvanicum gen. nov., sp. nov., a novel betaproteobacterium isolated from a waste-treating bioreactor.

A new betaproteobacterium, CGI-09T, was isolated from an activated sludge bioreactor which treated landfill leachate. Based on 16S rRNA gene sequence analysis, the new strain shared the highest pairwise similarity values with members of the order Burkholderiales: Derxia gummosa IAM 13946T (family Alcaligenaceae), 93.7 % and Lautropia mirabilis DSM 11362T (family Burkholderiaceae), 93.6 %. Cells of strain CGI-09T were rod-shaped and non-motile. The new strain was oxidase and catalase positive and capable of reducing nitrate to nitrite. The predominant fatty acids were C16 : 1 ω7c, C16 : 0, cycloC17 : 0 and C18 : 1 ω7c, the major respiratory quinone was Q-8, and the detected polar lipids were phosphatidylglycerol, diphosphatidylglycerol, phosphatidylethanolamine and an unknown phospholipid. The G+C content of the genomic DNA of strain CGI-09T was 70.2 mol%. The new bacterium can be distinguished from the members of genera Derxia and Lautropia based on its non-motile cells, arginine dihydrolase activity, its high cyclo C17 : 0 fatty acid content and the lack of hydroxy fatty acids. On the basis of the phenotypic, chemotaxonomic and molecular data, strain CGI-09T is considered to represent a new genus and species within the family Burkholderiaceae, for which the name Quisquiliibacterium transsilvanicum gen. nov., sp. nov. is proposed. The type strain is CGI-09T (=DSM 29781T=JCM 31785T).

Caenimicrobium hargitense gen. nov., sp. nov., a new member of the family Alcaligenaceae (Betaproteobacteria) isolated from activated sludge.

A new betaproteobacterium, CGII-59m2T, was isolated from an activated sludge bioreactor which treated landfill leachate. The 16S rRNA gene sequence analysis revealed that strain CGII-59m2T belonged to the family Alcaligenaceae and shared the highest pairwise similarity values with Parapusillimonas granuli LMG 24012T (97.7 %), various species of the genus Bordetella (97.3-97.0 %) and Candidimonas nitroreducens LMG 24812T (97.0 %). Cells of strain CGII-59m2T were rod-shaped, non-motile, and oxidase- and catalase-positive. The predominant fatty acids were C16 : 1ω7c, C16 : 0, cyclo C17 : 0 and C18 : 1ω7c, the major respiratory quinone was Q-8, and the main polar lipids were phosphatidylethanolamine, phosphatidylglycerol, diphosphatidylglycerol and an unknown phospholipid. The G+C content of the genomic DNA of strain CGII-59m2T was 62.3 mol%. The new bacterium can be distinguished from the closely related type strains based on its non-motile cells and its high C16 : 1ω7c fatty acid content. On the basis of the phenotypic, chemotaxonomic and molecular data, strain CGII-59m2T is considered to represent a novel species of a new genus, for which the name Caenimicrobium hargitense gen. nov., sp. nov. is proposed. The type strain is CGII-59m2T (=DSM 29806T=NCAIM B.02615T).

Rufibacter quisquiliarum sp. nov., a new member of the phylum Bacteroidetes isolated from a bioreactor treating landfill leachate.

A novel bacterium, CAI-18bT, was isolated from a bioreactor that treated landfill leachate using an oligotrophic growth medium. Phylogenetic analysis based on the 16S rRNA gene sequence revealed that strain CAI-18bT is a member of the genus Rufibacter, showing 97.1 % pairwise similarity to Rufibacter roseus H359T, 96.4 % to Rufibacter tibetensis 1351T, 96.4 % to Rufibacter glacialis MDT1-10-3T and 96.0 % to Rufibacter immobilis MCC P1T. Strain CAI-18bT was rod-shaped, motile, oxidase- and catalase-positive. The predominant fatty acids were iso-C15 : 0 (24.1 %) and iso-C17 : 1 I (22.3 %), the major respiratory quinone was MK-7, and the predominant polar lipids were phosphatidylethanolamine and an unknown aminophospholipid. The G+C content of the genomic DNA of strain CAI-18bT was 50.7 mol%. The novel bacterium can be distinguished from related type strains based on its ability to assimilate N-acetylglucosamine and gentiobiose. On the basis of the phenotypic, chemotaxonomic and molecular data, strain CAI-18bT represents a novel species of the genus Rufibacter, for which the name Rufibacter quisquiliarum sp. nov. is proposed. The type strain is CAI-18bT (=DSM 29854T=NCAIM B.02614T).

Unique picoeukaryotic algal community under multiple environmental stress conditions in a shallow, alkaline pan.

Winter phytoplankton communities in the shallow alkaline pans of Hungary are frequently dominated by picoeukaryotes, sometimes in particularly high abundance. In winter 2012, the ice-covered alkaline Zab-szék pan was found to be extraordinarily rich in picoeukaryotic green algae (42-82 × 10(6) cells ml(-1)) despite the simultaneous presence of multiple stressors (low temperature and light intensity with high pH and salinity). The maximum photosynthetic rate of the picoeukaryote community was 1.4 µg C µg chlorophyll a (-1) h(-1) at 125 µmol m(-2) s(-1). The assimilation rates compared with the available light intensity measured on the field show that the community was considerably light-limited. Estimated areal primary production was 180 mg C m(-2) d(-1). On the basis of the 18S rRNA gene analysis (cloning and DGGE), the community was phylogenetically heterogeneous with several previously undescribed chlorophyte lineages, which indicates the ability of picoeukaryotic communities to maintain high genetic diversity under extreme conditions.