Phylogenetic Profiles Research Articles

Virulence factors and phylogenetic group profile of uterine Escherichia coli in early postpartum of high-producing dairy cows

Escherichia coli is the most common contaminant of the bovine uterus in the first few weeks postpartum, and one of the most important pathogens involved in uterine infections. This bacterium is characterised by diverse virulence factors (VF); however, the profile of E. coli VF in physiologic postpartum uterine contamination is not well established. Therefore, the present study aimed to investigate the prevalence of intrauterine bacteria, a set of virulence factors and phylogroups of E. coli isolates, leukogram and uterine cytology in 75 Holstein cows at 24 h (Sampling time 1) and 14 days (Sampling time 2) postpartum. Escherichia coli, α-haemolytic Streptococcus, Trueperella pyogenes, and Pasteurella multocida were the most common microorganisms isolated in pure culture, whereas E. coli and Clostridium perfringens Type A, E. coli and α-haemolytic Streptococcus, and E. coli and Proteus mirabilis were the most frequent microorganisms in mixed colonies at both studied sampling times. In the 59 E. coli isolates after 24 h (n = 35) and 14 days (n = 24) postpartum, the genes detected for VF were fimH, iucD/aer, kps, hlyA, usp, vt1 and vt2. Most E. coli strains detected in both moments of study belonged to Commensal phylogenetic groups A and B1, whereas Pathogenic groups D and B2 were identified at 24 h and 14 days postpartum respectively. All cows presented suitable immune response against the presence of bacteria in uterine lumen, observed by leukocytosis, neutrophilia, lymphocytosis and monocytosis at leukogram and a high number of polymorphonuclear leukocytes in uterine cytology, in both studied moments. Thus, a complex diversity of aerobic and anaerobic bacteria is involved in uterine contamination in the early postpartum of cows, besides the predominance of E. coli. Moreover, the genes fimH, iucD/aer, hlyA, kps, usp, vt1 and vt2 play a key role in the virulence of E. coli in this period.

Dengue Virus in Traveler Returning to Japan from the Democratic Republic of the Congo, 2015.

Dengue fever (DF) is a mosquito-borne disease and a significant global public health problem. Although a few serological surveys in the literature suggest endemic DF in many parts of Africa, DF cases in these countries are generally underreported because of the lack of diagnostic testing and systematic surveillance; thus, little is known about the phylogenetic profile of circulating strains. In April 2015, DF was diagnosed in a Japanese national returning from the Democratic Republic of the Congo (DRC). Dengue virus 1 (DENV-1) RNA was detected in the patient's serum sample using real-time reverse transcription PCR. Phylogenetic analysis of the E gene revealed that the detected DENV-1 strain was classified as genotype V and was closely related, with 100% nucleotide identity, to the strain causing the 2013 DF epidemic in Angola, which is located directly south of the DRC. This is the first report to characterize the circulating DENV strain in the DRC, and the findings indicate that the DENV-1 strain causing the 2013 DF epidemic in Angola was also circulating in the DRC in 2015.

The Acinetobacter trimeric autotransporter adhesin Ata controls key virulence traits of Acinetobacter baumannii

ABSTRACTAcinetobacter baumannii is a Gram-negative pathogen that causes a multitude of nosocomial infections. The Acinetobacter trimeric autotransporter adhesin (Ata) belongs to the superfamily of trimeric autotransporter adhesins which are important virulence factors in many Gram-negative species. Phylogenetic profiling revealed that ata is present in 78% of all sequenced A. baumannii isolates but only in 2% of the closely related species A. calcoaceticus and A. pittii. Employing a markerless ata deletion mutant of A. baumannii ATCC 19606 we show that adhesion to and invasion into human endothelial and epithelial cells depend on Ata. Infection of primary human umbilical cord vein endothelial cells (HUVECs) with A. baumannii led to the secretion of interleukin (IL)-6 and IL-8 in a time- and Ata-dependent manner. Furthermore, infection of HUVECs by WT A. baumannii was associated with higher rates of apoptosis via activation of caspases-3 and caspase-7, but not necrosis, in comparison to ∆ata. Ata deletion mutants were furthermore attenuated in their ability to kill larvae of Galleria mellonella and to survive in larvae when injected at sublethal doses. This indicates that Ata is an important multifunctional virulence factor in A. baumannii that mediates adhesion and invasion, induces apoptosis and contributes to pathogenicity in vivo.

MYORG is associated with recessive primary familial brain calcification.

ObjectiveTo investigate the genetic basis of the recessive form of primary familial brain calcification and study pathways linking a novel gene with known dominant genes that cause the disease.MethodsWhole exome sequencing and Sanger‐based segregation analysis were used to identify possible disease causing mutations. Mutation pathogenicity was validated by structural protein modeling. Functional associations between the candidate gene, MYORG, and genes previously implicated in the disease were examined through phylogenetic profiling.ResultsWe studied nine affected individuals from two unrelated families of Middle Eastern origin. The median age of symptom onset was 29.5 years (range 21–57 years) and dysarthria was the most common presenting symptom. We identified in the MYORG gene, a homozygous c.1233delC mutation in one family and c.1060_1062delGAC mutation in another. The first mutation results in protein truncation and the second in deletion of a highly conserved aspartic acid that is likely to disrupt binding of the protein with its substrate. Phylogenetic profiling analysis of the MYORG protein sequence suggests co‐evolution with a number of calcium channels as well as other proteins related to regulation of anion transmembrane transport (False Discovery Rate, FDR < 10−8) and with PDCD6IP, a protein interacting with PDGFR β which is known to be involved in the disease.Interpretation MYORG mutations are linked to a recessive form of primary familial brain calcification. This association was recently described in patients of Chinese ancestry. We suggest the possibility that MYORG mutations lead to calcification in a PDGFR β‐related pathway.

OrthoInspector 3.0: open portal for comparative genomics.

OrthoInspector is one of the leading software suites for orthology relations inference. In this paper, we describe a major redesign of the OrthoInspector online resource along with a significant increase in the number of species: 4753 organisms are now covered across the three domains of life, making OrthoInspector the most exhaustive orthology resource to date in terms of covered species (excluding viruses). The new website integrates original data exploration and visualization tools in an ergonomic interface. Distributions of protein orthologs are represented by heatmaps summarizing their evolutionary histories, and proteins with similar profiles can be directly accessed. Two novel tools have been implemented for comparative genomics: a phylogenetic profile search that can be used to find proteins with a specific presence-absence profile and investigate their functions and, inversely, a GO profiling tool aimed at deciphering evolutionary histories of molecular functions, processes or cell components. In addition to the re-designed website, the OrthoInspector resource now provides a REST interface for programmatic access. OrthoInspector 3.0 is available at http://lbgi.fr/orthoinspectorv3.

Mangrove Sediment Microbiome: Adaptive Microbial Assemblages and Their Routed Biogeochemical Processes in Yunxiao Mangrove National Nature Reserve, China.

Microorganisms play important roles in mangrove ecosystems. However, we know little about the ecological implications of mangrove microbiomes for high productivity and the efficient circulation of elements in mangrove ecosystems. Here, we focused on mangrove sediments located at the Yunxiao National Mangrove Reserve in southeast China, uncovering the mangrove microbiome using the 16S rRNA gene and shotgun metagenome sequencing approaches. Physicochemical assays characterized the Yunxiao mangrove sediments as carbon (C)-rich, sulfur (S)-rich, and nitrogen (N)-limited environment. Then phylogenetic analysis profiling a distinctive microbiome with an unexpected high frequency of Chloroflexi and Nitrospirae appeared to be an adaptive characteristic of microbial structure in S-rich habitat. Metagenome sequencing analysis revealed that the metabolic pathways of N and S cycling at the community-level were routed through ammonification and dissimilatory nitrate reduction to ammonium for N conservation in this N-limited habitat, and dissimilatory sulfate reduction along with polysulfide formation for generating bioavailable S resource avoiding the biotoxicity of sulfide in mangrove sediments. In addition, methane metabolism acted as a bridge to connect C cycling to N and S cycling. Further identification of possible biogeochemical linkers suggested Syntrophobacter, Sulfurovum, Nitrospira, and Anaerolinea potentially drive the coupling of C, N, and S cycling. These results highlighting the adaptive routed metabolism flow, a previously undescribed property of mangrove sediment microbiome, appears to be a defining characteristic of this habitat and may significantly contribute to the high productivity of mangrove ecosystems, which could be used as indicators for the health and biodiversity of mangrove ecosystems.

Phylogenetic, toxigenic and virulence profiles of Alternaria species causing leaf blight of tomato in Egypt

Species of Alternaria are serious plant pathogens, causing major losses on a wide range of crops. Leaf blight symptoms were observed on tomato leaves, and samples were collected from various regions. Isolation was done from symptomatic tomato leaves, and 15 representatives were selected from a collection of 65 isolates of Alternaria species. The virulence of Alternaria isolates was investigated on detached leaves (DL) and whole plants of tomato cv. Super strain B. A phylogenetic analysis was performed based on three partial gene regions, the glyceraldehyde-3-phosphate dehydrogenase (GAPDH), the RNA polymerase second largest subunit (RPB2) and the Alternaria major allergen gene (Alt a 1). The potentiality of Alternaria isolates to produce toxins was also investigated on the basis of thin-layer chromatography (TLC). Our investigations revealed that Alternaria isolates showed different levels of virulence either on tomato plants or DL. Based on the phylogeny of three genes, Alternaria isolates encompassed two species of small-spored morphospecies: A. alternata (14 isolates) and A. arborescens (single isolate). The produced toxins varied among Alternaria isolates with tenuazonic acid (TeA) being the most abundant mycotoxin produced by most isolates. This study highlighted on other Alternaria species in Egypt that might represent a serious concern for tomato producers as causal agents of leaf blight over other species, i.e. A. solani.

Abstract IA14: Circulating tumor DNA in early-stage NSCLC: A lung TRACERx study

Abstract The Cancer Research UK-funded study TRACERx (TRAcking Cancer Evolution through therapy [Rx]) aims to recruit patients undergoing surgery for early-stage non-small cell lung cancer (NSCLC). Patients donate blood pre- and postoperatively alongside multi-region sampled primary tumor tissue. Up to 40% of patients undergoing surgery for NSCLC experience relapse of their disease. At this point patients are approached for relapse tissue sampling. TRACERx therefore provides a strong platform for evaluating circulating tumor DNA (ctDNA) in early-stage NSCLC and a minimal residual disease (MRD) setting. In this presentation we will describe data arising from a phylogenetic, patient-specific approach to ctDNA evaluation in the first 100 patients analyzed as part of TRACERx. We will discuss clinicopathologic factors associated with ctDNA detection in early-stage NSCLC and suggest the existence of distinct tumor phenotypic characteristics that predict the presence of ctDNA at measurable quantities in plasma. Additionally, we will discuss tumor volume limit of detection analyses and highlight potential implications of our findings within the context of early cancer detection. We will then examine data arising from the ctDNA profiling of postoperative plasma samples taken in the adjuvant setting. These data demonstrate that ctDNA detection is specific for NSCLC relapse and that ctDNA detection precedes clinical diagnosis of NSCLC relapse (median lead-time of 77 days, range 10 to 346 days). We will provide examples of using phylogenetic ctDNA profiling to characterize the subclonal nature of relapsing NSCLC as a potential method to personalize relapse therapeutic intervention. Finally, we will highlight potential advantages of patient-specific ctDNA profiling in an adjuvant MRD setting and suggest ways that this technology can be used to fuel adjuvant drug development in NSCLC by driving novel study designs. Citation Format: Christopher Abbosh. Circulating tumor DNA in early-stage NSCLC: A lung TRACERx study [abstract]. In: Proceedings of the Fifth AACR-IASLC International Joint Conference: Lung Cancer Translational Science from the Bench to the Clinic; Jan 8-11, 2018; San Diego, CA. Philadelphia (PA): AACR; Clin Cancer Res 2018;24(17_Suppl):Abstract nr IA14.

Rubella Virus Genotype 1E in Travelers Returning to Japan from Indonesia, 2017

Although rubella is epidemic in Indonesia, the phylogenetic profile of circulating rubella virus strains has not been clarified. In 2017, rubella virus was detected in 2 travelers who returned from Indonesia to Japan. These strains were classified into genotype 1E lineage 2, which may be an indigenous strain in Indonesia.

Schlafen2 mutation in mice causes an osteopetrotic phenotype due to a decrease in the number of osteoclast progenitors

Osteoclasts are the bone resorbing cells that derive from myeloid progenitor cells. Although there have been recent advancements in the ability to identify osteoclast progenitors, very little is known about the molecular mechanisms governing their homeostasis. Here, by analyzing the normalized phylogenetic profiles of the Schlafen (Slfn) gene family, we found that it co-evolved with osteoclast-related genes. Following these findings, we used a Slfn2 loss-of-function mutant mouse, elektra, to study the direct role of Slfn2 in osteoclast development and function. Slfn2eka/eka mice exhibited a profound increase in their cancellous bone mass and a significant reduction in osteoclast numbers. In addition, monocyte cultures from the bone marrow of Slfn2eka/eka mice showed a reduction in osteoclast number and total resorption area. Finally, we show that the bone marrow of Slfn2eka/eka mice have significantly less CD11b–Ly6Chi osteoclast precursors. Overall, our data suggest that Slfn2 is required for normal osteoclast differentiation and that loss of its function in mice results in an osteopetrotic phenotype.

Phylogenetic Clustering of Genes Reveals Shared Evolutionary Trajectories and Putative Gene Functions.

Homologous genes in prokaryotes can be described using phylogenetic profiles which summarize their patterns of presence or absence across a set of genomes. Phylogenetic profiles have been used for nearly twenty years to cluster genes based on measures such as the Euclidean distance between profile vectors. However, most approaches do not take into account the phylogenetic relationships amongst the profiled genomes, and overrepresentation of certain taxonomic groups (i.e., pathogenic species with many sequenced representatives) can skew the interpretation of profiles. We propose a new approach that uses a coevolutionary method defined by Pagel to account for the phylogenetic relationships amongst target organisms, and a hierarchical-clustering approach to define sets of genes with common distributions across the organisms. The clusters we obtain using our method show greater evidence of phylogenetic and functional clustering than a recently published approach based on hidden Markov models. Our clustering method identifies sets of amino-acid biosynthesis genes that constitute cohesive pathways, and motility/chemotaxis genes with common histories of descent and lateral gene transfer.

Phylogenetic analysis, biofilm production, and antimicrobial resistance profile of Escherichia coli isolated from slaughtered pigs

ABSTRACT Enteric diseases of bacterial origin are frequent in the pig industry, of particular notoriety are the colibacillosis that mainly affect piglets and cause great damage to the swine industry worldwide. The aim of the study was to analyze phylogenetics, to detect biofilm production, and to determine antimicrobial resistance profile in 126 strains of Escherichia coli isolated from swabs obtained from fragments of the small intestines of 235 healthy pigs killed in slaughterhouses in Pernambuco (Brazil) using polymerase chain reaction (PCR), adherence to microplates test and disc diffusion technique. Of the analyzed samples, 88.10% (111/126) were classified in phylogenetic group B1; 4.76% (6/126) in group D; 3.97% (5/126) in group B2 and, 3.17% (4/126) in group A. Antimicrobial resistance rates observed were: lincomycin 100% (126/126), erythromycin 100% (126/126), chlortetracycline 94.44% (119/126), cephalothin 51.59% (65/126), ampicillin 38.89% (49/126), sulfamethoxazole + trimethoprim 37.3% (47/126), ciprofloxacin 19.84% (25/126), norfloxacin 14.29% (18/126), gentamicin 8.73% (11/126) and, chloramphenicol 5.55% (7/126). Multiple antibiotic resistance (MAR) ranged from 0.2 to 0.9. Of the strains tested 46.03% (58/126) produced biofilm, and 99.21% (125/126) of the strains exhibited multi-resistance. Further studies are required to elucidate the importance of each phylogenetic group in pigs and to prevent the propagation of multi-resistant E. coli strains.

Genome-wide identification, and phylogenetic and expression profiling analyses of CaM and CML genes in Brassica rapa and Brassica oleracea

Calmodulin (CaM) and calmodulin-like (CML) proteins are two kinds of calcium (Ca2+)-sensing proteins that are involved in Ca2+-signaling processes. B. rapa and B. oleracea are two of three diploid Brassica species that have undergone recent additional Brassiceae-lineage-specific whole-genome triplication events. To elucidate the expansion, evolution, expression pattern and interaction network of these Ca2+ sensors in B. rapa and B. oleracea, we conducted a comparative syntenic study on a genome-wide level using Arabidopsis thaliana. In total, 80 and 79 CaM/CML genes were identified in B. rapa and B. oleracea, respectively. The CaM/CML genes have expanded throughout the whole genomes of B. rapa and B. oleracea by whole-genome and tandem duplication events. The CaMs/CMLs of B. rapa and B. oleracea can be classified into nine subgroups based on their A. thaliana orthologs. Expression data from various tissues revealed that a large portion of CaM/CML genes exhibited patterns of differential and tissue-specific expression. The expression levels among duplicated paralogs in different subgenomes demonstrated the expression divergence of these genes in B. rapa and B. oleracea. The differential expressions of duplicated CaM and CML genes in B. rapa indicated that their functional differentiation occurred after polyploidization. The construction of interaction network and GO enrichment analysis of genes in this network revealed Br/BoCaMs/CMLs involving biological processes. This work will promote a better understanding of Ca2+ sensors and Ca2+-signaling pathways in B. rapa and B. oleracea.

Metagenomic insights into the phylogenetic and functional profiles of soil microbiome from a managed mangrove in Malaysia

The Matang Mangrove Forest Reserve (MMFR) in Malaysia has been acknowledged as the best sustainably-managed mangrove forest in the world. The management employed a 30-year rotation cycle to enable harvest of mangrove timbers, followed by replanting and series of thinnings. Information on soil microbial diversity and its functional capability in managed mangrove is scarce. To this end, we analysed high-throughput metagenomic datasets from two sites at MMFR with distinct features: the Virgin Jungle Forest and the harvested Productive Zone. Taxonomic classification of sequencing reads using metagenomics RAST (MG-RAST) revealed distinct differences at phylum and class level for bacteria present in both samples. Productive zone, which consisted of managed forest blocks, exhibited total organic carbon content that was 18.7 times higher than that of Virgin Jungle Forest. The Productive Zone microbial community was overabundant in genes related to carbohydrate metabolism, especially enzymes involved in the degradation and utilization of polysaccharides from plant cell wall. Functional analysis focusing on carbohydrate degrading enzymes revealed an array of enzymes involved in hemicellulose, cellulose and pectin utilization enzymes in Matang mangrove metagenomes. Overall, differences in taxonomic composition and function profile of soil microorganisms were observed between Productive Zone and Virgin Jungle Forest. The results presented in this study have important implications in understanding managed mangrove microbial assemblage and provide important resources for the discovery of bacterial species and enzymes involved in litter processing.

The evolutionary signal in metagenome phyletic profiles predicts many gene functions

BackgroundThe function of many genes is still not known even in model organisms. An increasing availability of microbiome DNA sequencing data provides an opportunity to infer gene function in a systematic manner.ResultsWe evaluated if the evolutionary signal contained in metagenome phyletic profiles (MPP) is predictive of a broad array of gene functions. The MPPs are an encoding of environmental DNA sequencing data that consists of relative abundances of gene families across metagenomes. We find that such MPPs can accurately predict 826 Gene Ontology functional categories, while drawing on human gut microbiomes, ocean metagenomes, and DNA sequences from various other engineered and natural environments. Overall, in this task, the MPPs are highly accurate, and moreover they provide coverage for a set of Gene Ontology terms largely complementary to standard phylogenetic profiles, derived from fully sequenced genomes. We also find that metagenomes approximated from taxon relative abundance obtained via 16S rRNA gene sequencing may provide surprisingly useful predictive models. Crucially, the MPPs derived from different types of environments can infer distinct, non-overlapping sets of gene functions and therefore complement each other. Consistently, simulations on > 5000 metagenomes indicate that the amount of data is not in itself critical for maximizing predictive accuracy, while the diversity of sampled environments appears to be the critical factor for obtaining robust models.ConclusionsIn past work, metagenomics has provided invaluable insight into ecology of various habitats, into diversity of microbial life and also into human health and disease mechanisms. We propose that environmental DNA sequencing additionally constitutes a useful tool to predict biological roles of genes, yielding inferences out of reach for existing comparative genomics approaches.

Phylogeny and divergence times of suckers (Cypriniformes: Catostomidae) inferred from Bayesian total-evidence analyses of molecules, morphology, and fossils.

Catostomidae (“suckers”) is a diverse (76 species) and broadly distributed family of Holarctic freshwater fishes with a rich fossil record and a considerable number (∼35%) of threatened and imperiled species. We integrate DNA sequences (three mitochondrial genes, three nuclear genes), morphological data, and fossil information to infer sucker phylogenetic relationships and divergence times using Bayesian “total-evidence” methods, and then test hypotheses about the temporal diversification of the group. Our analyses resolved many nodes within subfamilies and clarified Catostominae relationships to be of the form ((Thoburniini, Moxostomatini), (Erimyzonini, Catostomini)). Patterns of subfamily relationships were incongruent, but mainly supported two placements of the Myxocyprininae; distinguishing these using Bayes factors lent strongest support to a model with Myxocyprininae sister to all remaining sucker lineages. We improved our Bayesian total-evidence dating analysis by excluding problematic characters, using a clock-partitioning scheme identified by Bayesian model selection, and employing a fossilized birth-death tree prior accommodating morphological data and fossils. The resulting chronogram showed that suckers evolved since the Late Cretaceous–Eocene, and that the Catostomini and Moxostomatini clades have accumulated species diversity since the early to mid-Miocene. These results agree with the fossil record and confirm previous hypotheses about dates for the origins of Catostomide and catostomine diversification, but reject previous molecular hypotheses about the timing of divergence of ictiobines, and between Asian–North American lineages. Overall, our findings from a synthesis of multiple data types enhance understanding of the phylogenetic relationships, taxonomic classification, and temporal diversification of suckers, while also highlighting practical methods for improving Bayesian divergence dating models by coupling phylogenetic informativeness profiling with relaxed-clock partitioning.

Taxonomy-aware feature engineering for microbiome classification

BackgroundWhat is a healthy microbiome? The pursuit of this and many related questions, especially in light of the recently recognized microbial component in a wide range of diseases has sparked a surge in metagenomic studies. They are often not simply attributable to a single pathogen but rather are the result of complex ecological processes. Relatedly, the increasing DNA sequencing depth and number of samples in metagenomic case-control studies enabled the applicability of powerful statistical methods, e.g. Machine Learning approaches. For the latter, the feature space is typically shaped by the relative abundances of operational taxonomic units, as determined by cost-effective phylogenetic marker gene profiles. While a substantial body of microbiome/microbiota research involves unsupervised and supervised Machine Learning, very little attention has been put on feature selection and engineering.ResultsWe here propose the first algorithm to exploit phylogenetic hierarchy (i.e. an all-encompassing taxonomy) in feature engineering for microbiota classification. The rationale is to exploit the often mono- or oligophyletic distribution of relevant (but hidden) traits by virtue of taxonomic abstraction. The algorithm is embedded in a comprehensive microbiota classification pipeline, which we applied to a diverse range of datasets, distinguishing healthy from diseased microbiota samples.ConclusionWe demonstrate substantial improvements over the state-of-the-art microbiota classification tools in terms of classification accuracy, regardless of the actual Machine Learning technique while using drastically reduced feature spaces. Moreover, generalized features bear great explanatory value: they provide a concise description of conditions and thus help to provide pathophysiological insights. Indeed, the automatically and reproducibly derived features are consistent with previously published domain expert analyses.

PROBE: analysis and visualization of protein block-level evolution.

Comparative studies of protein sequences are widely used in evolutionary and comparative genomics studies, but there is a lack of efficient tools to identify conserved regions ab initio within a protein multiple alignment. PROBE provides a fully automatic analysis of protein family conservation, to identify conserved regions, or 'blocks', that may correspond to structural/functional domains or motifs. Conserved blocks are identified at two different levels: (i) family level blocks indicate sites that are probably of central importance to the protein's structure or function, and (ii) sub-family level blocks highlight regions that may signify functional specialization, such as binding partners, etc. All conserved blocks are mapped onto a phylogenetic tree and can also be visualized in the context of the multiple sequence alignment. PROBE thus facilitates in-depth studies of sequence-structure-function-evolution relationships, and opens the way to block-level phylogenetic profiling. Freely available on the web at http://www.lbgi.fr/∼julie/probe/web.

Prediction of Protein-Protein Interactions Using An Effective Sequence Based Combined Method

Abstract Proteins and their interactions play a key role in the realization of all cellular biological activities of organisms. Therefore, prediction of protein-protein interactions is crucial for elucidating biological processes. Experimental studies are inadequate for some reasons such as the time required to reveal interactions, the fact that it is an expensive way and the number of yet unknown interactions is too great. Thus, a number of computational methods have been developed to predict protein-protein interactions. Generally, many of these methods that produce good results cannot be used without additional biological information such as protein domains, protein structural information, gene neighborhoods, gene expressions, and phylogenetic profiles. Therefore, there is a need for computational methods that can successfully predict interactions using only protein sequences. In this study, we present a novel sequence-based computational model. We applied a new technique called weighted skip-sequential conjoint triads in the proposed method. The results of this research were evaluated on generally used databases and demonstrated its success in this field.

The surveillance of pre-mRNA splicing is an early step in C. elegans RNAi of endogenous genes.

RNAi pathways detect and silence foreign nucleic acids such as viruses as well as endogenous genes in many species. The phylogenetic profile across eukaryotes of proteins that mediate key steps in RNAi is correlated with the profiles of multiple mRNA splicing proteins and with intron number, suggesting that RNAi may surveil mRNA splicing to detect the divergent or absent introns of viruses. Here we examine the role of mRNA splicing in Caenorhabditis elegans RNAi. We found that viable null mutations in U1 and U2 small nuclear ribonucleic protein (snRNP)-specific splicing factor genes cause defects in RNAi. The U1A ortholog rnp-2 is required for normal ERGO-1 Argonaute class 26G siRNA biogenesis, trans-splicing of the eri-6/7 transcript, and targeting of poorly conserved gene transcripts by WAGO Argonaute class 22G siRNAs. We found that gene transcripts engaged by the siRNA-generating machinery are poorly conserved, possess few introns, and often have introns that are divergent from introns with strong consensus splicing sites found in highly conserved genes. We present biochemical evidence that RNAi targeted transcripts are tightly bound to spliceosomes. These findings suggest multiple layers of regulation by the spliceosome at early steps of small RNA-mediated gene silencing.