A Study of Horizontally Transferred Glycosyl Hydrolase Family 6 Genes in Tunicate Genomes

Tunicates are the closest extant relatives of vertebrates. Tunicates produce cellulosecontaining tunic and exhibit very characteristic lifestyles among animals. Their unique ability to synthesize cellulose results from a horizontally transferred cellulose synthase gene (CesA). Interestingly, a Glycosyl Hydrolase Family 6 (GH6) hydrolase-like domain exists at the C-terminus of tunicate CesA but not in cellulose synthases of other organisms. This led to the identification of another independent GH6-encoding gene, GH6-1, in tunicate genomes. These GH6-encoding genes exist exclusively in tunicates within the animal kingdom. The existence of GH6-encoding genes and the combination of GH6 and cellulose synthase domains raised the question of the evolutionary origin and function of GH6s in tunicates. To answer these questions, I first examined the phylogenetic relationship of GH6- encoding genes by comparing their sequence signatures. Tunicate CesA and GH6-1 genes represent two independent orthologous groups, but the origin of these genes before a horizontal transfer event could not be ascertained. Secondly, I examined the expression of tunicate CesA and GH6-1 genes in Ciona intestinalis type A, a model ascidian tunicate. The gene expression in embryos at early developmental stages was examined by quantitative reverse transcription PCR and in situ hybridization. Obvious expression of both CesA and GH6-1 were found at embryonic stages of Ciona embryo at epidermis. The observed expression profiles were also compared with a set of single-cell transcriptome data provided by our collaborators. Embryonic cells of late tailbud stage I showed that both GH6-1- and CesA-expressing cells are mostly in cell clusters of epidermal identity. Localized signal in the reporter assay also suggest the existence of specific enhancers upstream to Ciona GH6- 1 gene. Finally, I used genome-editing technique to generate GH6-1 knockout larvae of C. intestinalis type A and observed that affected embryos show perturbed papillae formation and metamorphosis. My study showed that GH6-1, a gene very likely originated from horizontal gene transfer, is recruited to function in ascidian early development. This observation would help to address how tunicates evolved by obtaining their unique anatomy and ecology.Okinawa Institute of Science and Technology Graduate Universit

Genome-wide survey and expression analysis of the bHLH-PAS genes in the amphioxus Branchiostoma floridae reveal both conserved and diverged expression patterns between cephalochordates and vertebrates

BACKGROUND: The bHLH-PAS transcription factors are found in both protostomes and deuterostomes. They are involved in many developmental and physiological processes, including regional differentiation of the central nervous system, tube-formation, hypoxia signaling, aromatic hydrocarbon sensing, and circadian rhythm regulation. To understand the evolution of these genes in chordates, we analyzed the bHLH-PAS genes of the basal chordate amphioxus (Branchiostoma floridae). RESULTS: From the amphioxus draft genome database, we identified ten bHLH-PAS genes, nine of which could be assigned to known orthologous families. The tenth bHLH-PAS gene could not be assigned confidently to any known bHLH family; however, phylogenetic analysis clustered this gene with arthropod Met family genes and two spiralian bHLH-PAS-containing sequences, suggesting that they may share the same ancestry. We examined temporal and spatial expression patterns of these bHLH-PAS genes in developing amphioxus embryos. We found that BfArnt, BfNcoa, BfSim, and BfHifα were expressed in the central nervous system in patterns similar to those of their vertebrate homologs, suggesting that their functions may be conserved. By contrast, the amphioxus BfAhr and BfNpas4 had expression patterns distinct from those in vertebrates. These results imply that there were changes in gene regulation after the divergence of cephalochordates and vertebrates. CONCLUSIONS: We have identified ten bHLH-PAS genes from the amphioxus genome and determined the embryonic expression profiles for these genes. In addition to the nine currently recognized bHLH-PAS families, our survey suggests that the BfbHLHPAS-orphan gene along with arthropod Met genes and the newly identified spiralian bHLH-PAS-containing sequences represent an ancient group of genes that were lost in the vertebrate lineage. In a comparison with the expression patterns of the vertebrate bHLH-PAS paralogs, which are the result of whole-genome duplication, we found that although several members seem to retain conserved expression patterns during chordate evolution, many duplicated paralogs may have undergone subfunctionalization and neofunctionalization in the vertebrate lineage. In addition, our survey of amphioxus bHLH-PAS gene models from genome browser with experimentally verified cDNA sequences calls into question the accuracy of the current in silico gene annotation of the B. floridae genome

Generalizable Resource Allocation in Stream Processing via Deep Reinforcement Learning

This paper considers the problem of resource allocation in stream processing, where continuous data flows must be processed in real time in a large distributed system. To maximize system throughput, the resource allocation strategy that partitions the computation tasks of a stream processing graph onto computing devices must simultaneously balance workload distribution and minimize communication. Since this problem of graph partitioning is known to be NP-complete yet crucial to practical streaming systems, many heuristic-based algorithms have been developed to find reasonably good solutions. In this paper, we present a graph-aware encoder-decoder framework to learn a generalizable resource allocation strategy that can properly distribute computation tasks of stream processing graphs unobserved from training data. We, for the first time, propose to leverage graph embedding to learn the structural information of the stream processing graphs. Jointly trained with the graph-aware decoder using deep reinforcement learning, our approach can effectively find optimized solutions for unseen graphs. Our experiments show that the proposed model outperforms both METIS, a state-of-the-art graph partitioning algorithm, and an LSTM-based encoder-decoder model, in about 70% of the test cases.Comment: Accepted by AAAI 202

Phylogenetic Analyses of Glycosyl Hydrolase Family 6 Genes in Tunicates: Possible Horizontal Transfer

Horizontal gene transfer (HGT) is the movement of genetic material between different species. Although HGT is less frequent in eukaryotes than in bacteria, several instances of HGT have apparently shaped animal evolution. One well-known example is the tunicate cellulose synthase gene, CesA, in which a gene, probably transferred from bacteria, greatly impacted tunicate evolution. A Glycosyl Hydrolase Family 6 (GH6) hydrolase-like domain exists at the C-terminus of tunicate CesA, but not in cellulose synthases of other organisms. The recent discovery of another GH6 hydrolase-like gene (GH6-1) in tunicate genomes further raises the question of how tunicates acquired GH6. To examine the probable origin of these genes, we analyzed the phylogenetic relationship of GH6 proteins in tunicates and other organisms. Our analyses show that tunicate GH6s, the GH6-1 gene, and the GH6 part of the CesA gene, form two independent, monophyletic gene groups. We also compared their sequence signatures and exon splice sites. All tunicate species examined have shared splice sites in GH6-containing genes, implying ancient intron acquisitions. It is likely that the tunicate CesA and GH6-1 genes existed in the common ancestor of all extant tunicates

Improved sample dispersion in cryo-EM using “perpetually-hydrated” graphene oxide flakes

For many macromolecular complexes, the inability to uniformly disperse solubilized specimen particles within vitreous ice films precludes their analysis by cryo-electron microscopy (cryo-EM). Here, we introduce a sample preparation process using “perpetually-hydrated” graphene oxide flakes as particle support films, and report vastly improved specimen dispersion. The new method introduced in this study incorporates hydrated graphene oxide flakes into a standard sample preparation regime, without the need for additional tools or devices, making it a cost-effective and easily adoptable alternative to currently available sample preparation approaches

Molecular signature of clinical severity in recovering patients with severe acute respiratory syndrome coronavirus (SARS-CoV)

BACKGROUND: Severe acute respiratory syndrome (SARS), a recent epidemic human disease, is caused by a novel coronavirus (SARS-CoV). First reported in Asia, SARS quickly spread worldwide through international travelling. As of July 2003, the World Health Organization reported a total of 8,437 people afflicted with SARS with a 9.6% mortality rate. Although immunopathological damages may account for the severity of respiratory distress, little is known about how the genome-wide gene expression of the host changes under the attack of SARS-CoV. RESULTS: Based on changes in gene expression of peripheral blood, we identified 52 signature genes that accurately discriminated acute SARS patients from non-SARS controls. While a general suppression of gene expression predominated in SARS-infected blood, several genes including those involved in innate immunity, such as defensins and eosinophil-derived neurotoxin, were upregulated. Instead of employing clustering methods, we ranked the severity of recovering SARS patients by generalized associate plots (GAP) according to the expression profiles of 52 signature genes. Through this method, we discovered a smooth transition pattern of severity from normal controls to acute SARS patients. The rank of SARS severity was significantly correlated with the recovery period (in days) and with the clinical pulmonary infection score. CONCLUSION: The use of the GAP approach has proved useful in analyzing the complexity and continuity of biological systems. The severity rank derived from the global expression profile of significantly regulated genes in patients may be useful for further elucidating the pathophysiology of their disease

Selective predisposition to bacterial infections in IRAK-4–deficient children: IRAK-4–dependent TLRs are otherwise redundant in protective immunity

Human interleukin (IL) 1 receptor–associated kinase 4 (IRAK-4) deficiency is a recently discovered primary immunodeficiency that impairs Toll/IL-1R immunity, except for the Toll-like receptor (TLR) 3– and TLR4–interferon (IFN)-a/b pathways. The clinical and immunological phenotype remains largely unknown. We diagnosed up to 28 patients with IRAK-4 deficiency, tested blood TLR responses for individual leukocyte subsets, and TLR responses for multiple cytokines. The patients' peripheral blood mononuclear cells (PBMCs) did not induce the 11 non-IFN cytokines tested upon activation with TLR agonists other than the nonspecific TLR3 agonist poly(I:C). The patients' individual cell subsets from both myeloid (granulocytes, monocytes, monocyte-derived dendritic cells [MDDCs], myeloid DCs [MDCs], and plasmacytoid DCs) and lymphoid (B, T, and NK cells) lineages did not respond to the TLR agonists that stimulated control cells, with the exception of residual responses to poly(I:C) and lipopolysaccharide in MDCs and MDDCs. Most patients (22 out of 28; 79%) suffered from invasive pneumococcal disease, which was often recurrent (13 out of 22; 59%). Other infections were rare, with the exception of severe staphylococcal disease (9 out of 28; 32%). Almost half of the patients died (12 out of 28; 43%). No death and no invasive infection occurred in patients older than 8 and 14 yr, respectively. The IRAK-4–dependent TLRs and IL-1Rs are therefore vital for childhood immunity to pyogenic bacteria, particularly Streptococcus pneumoniae. Conversely, IRAK-4–dependent human TLRs appear to play a redundant role in protective immunity to most infections, at most limited to childhood immunity to some pyogenic bacteria

Changes in Corneal Basal Epithelial Phenotypes in an Altered Basement Membrane

To examine the corneal epithelial phenotype in an altered basement membrane.Corneas from 9 patients with symptoms of continuous unstable corneal curvature (CUCC) were harvested by penetrating keratoplasty and subjected to histology examination and immunohistochemical staining with transactivating and N-terminally truncated pP63 transcript (ΔNp63), cytokeratin 3 (Krt3), ATP-binding cassette sub-family G member 2 (ABCG2), connexin 43 (CX43), p38 mitogen-activated protein kinases (p38MAPK), activating protein 2 (TFAP2), and extracellular signal-regulated kinase (Erk1/2) monoclonal antibodies. Positive immunostaining with ABCG2, p38MAPK, and TFAP2 monoclonal antibodies was observed in the basal epithelial cells of CUCC patients, and CX43 and ΔNp63 were detected in the full-thickness epithelial cells of CUCC patients.Our results indicate that alteration of the corneal basement membrane induces a de-differentiation-like phenotype in corneal basal epithelial cells

Tree-Encoded Bitmaps

We propose a novel method to represent compressed bitmaps. Similarly to existing bitmap compression schemes, we exploit the compression potential of bitmaps populated with consecutive identical bits, i.e., 0-runs and 1-runs. But in contrast to prior work, our approach employs a binary tree structure to represent runs of various lengths. Leaf nodes in the upper tree levels thereby represent longer runs, and vice versa. The tree-based representation results in high compression ratios and enables efficient random access, which in turn allows for the fast intersection of bitmaps. Our experimental analysis with randomly generated bitmaps shows that our approach significantly improves over state-of-the-art compression techniques when bitmaps are dense and/or only barely clustered. Further, we evaluate our approach with real-world data sets, showing that our tree-encoded bitmaps can save up to one third of the space over existing techniques