Biology subjects

Zhang, R.

Publications and source records attributed to Zhang, R..

At least 613 records · Page 34Linked to original sources

bioRxiv2018

Single tube bead-based DNA co-barcoding for cost effective and accurate sequencing, haplotyping, and assembly

Obtaining accurate sequences from long DNA molecules is very important for genome assembly and other applications. Here we describe single tube long fragment read (stLFR), a technology that enables this a low cost. It is based on adding the same barcode sequence to sub-fragments of the original long DNA molecule (DNA co-barcoding). To achieve this efficiently, stLFR uses the surface of microbeads to create millions of miniaturized barcoding reactions in a single tube. Using a combinatorial process up to 3.6 billion unique barcode sequences were generated on beads, enabling practically non-redundant co-barcoding with 50 million barcodes per sample. Using stLFR, we demonstrate efficient unique co-barcoding of over 8 million 20-300 kb genomic DNA fragments. Analysis of the genome of the human genome NA12878 with stLFR demonstrated high quality variant calling and phasing into contigs up to N50 34 Mb. We also demonstrate detection of complex structural variants and complete diploid de novo assembly of NA12878. These analyses were all performed using single stLFR libraries and their construction did not significantly add to the time or cost of whole genome sequencing (WGS) library preparation. stLFR represents an easily automatable solution that enables high quality sequencing, phasing, SV detection, scaffolding, cost-effective diploid de novo genome assembly, and other long DNA sequencing applications.

genomics↗

bioRxiv2018

Inward tubulation of the plasma membrane expedites membrane exchange and receptor presentation

The plasma membrane is a crucial barrier between the cell and its external environment, and it also enables efficient passage of materials and information. Membrane vesicle trafficking allows precise delivery of materials but is rather inefficient. The mechanism for efficient membrane exchange remains elusive. Here we describe inward tubulation of the plasma membrane (PM tubes) that extends deep into the cytoplasm. These widespread PM tubes elongate along microtubules and are stabilized by actin filaments and cholesterol. PM tubes are preferred sites for connection between the endoplasmic reticulum and the plasma membrane. PM tubes facilitate receptor presentation at the surface of cells, possibly also shortening the distance for transported cargo to reach the external environment.\n\nIn BriefA new type of tubular membrane structures was discovered in cells, revealing a shortcut that cells employ to expedite material exchange with their external environment.\n\nHighlightsO_LIInward tubulation of the plasma membrane (PM tubes), transiently interacts with the Golgi apparatus\nC_LIO_LIMicrotubule side-binding proteins pull PM tubes, while actin filaments and cholesterol stabilize PM tubes\nC_LIO_LIPM tubes are preferred sites where ER-PM contacts form in response to increased cytoplasmic calcium concentration\nC_LIO_LIPM tubes are preferred sites for the surface presentation of GLUT1 upon glucose deprivation\nC_LI

cell biology↗

bioRxiv2018

Nonmuscle myosin II shRNA inhibit migration and contraction in rat hepatic stellate cells through regulating AKT/mTOR/S6K/4EBP1 signaling pathway

Migration and contraction of activated hepatic stellate cell (HSC) are essential factors for cirrhosis formation and development. It has been demonstrated that blebbistatin, a nonmuscle myosin II (NMMII) inhibitor, can inhibit the migration and contraction of HSC, whereas the main cell signaling pathway is still unknown. Mammalian target of rapamycin (mTOR) signaling pathway may be involved in many cells migration and contraction, whether NMMII and mTOR have any crosslinks draw our attention. In the currently study, we used LV-RNAi to specifically attenuate mTOR and NMMII in rat HSC. We aimed to examine the effect of mTOR LV-RNAi on the migration and contraction of HSC and explore the crosslink between mTOR cell signal and NMMII. Using real-time PCR and western blot, we found that mTOR and the downstream factors including S6K and 4EBP1 all up-regulated with the activation of HSC, mTOR and NMMII LV-RNAi was transfected into activated HSC using lipofectamine 2000. The levels of mRNA and proteins were also examined using real-time PCR and western blot respectively. The expression of mTOR can be down-regulated by NMMII LV-RNAi significantly, as well as the expression of S6K, 4EBP1, -SMA and collagen I, but the level of AKT was up-regulated. Then we used Transwell system and collagen lattices to examine the NMMII and mTOR LV-RNAi efficiency on HSC migration and contraction, as we hypothesized, both of the LV-RNAi could inhibit HSC migration and contraction significantly. These results indicated that nonmuscle myosin II shRNA inhibit migration and contraction in rat hepatic stellate cells through the regulation of mTOR/S6K/4EBP1 signaling pathway

cell biology↗

bioRxiv2018

Negative Cooperativity between Gemin2 and RNA Determines RNA Selection and Release of the SMN Complex in snRNP Assembly

The assembly of snRNP cores, in which seven Sm proteins, D1/D2/F/E/G/D3/B, form a ring around the nonameric Sm site of snRNAs, is the early step of spliceosome formation and essential to eukaryotes. It is mediated by the PMRT5 and SMN complexes sequentially in vivo. SMN deficiency causes neurodegenerative disease spinal muscular atrophy (SMA). How the SMN complex assembles snRNP cores is largely unknown, especially how the SMN complex achieves high RNA assembly specificity and how it is released. Here we show, using crystallographic and biochemical approaches, that Gemin2 of the SMN complex enhances RNA specificity of SmD1/D2/F/E/G via a negative cooperativity between Gemin2 and RNA in binding SmD1/D2/F/E/G. Gemin2, independent of its N-tail, constrains the horseshoe-shaped SmD1/D2/F/E/G from outside in a physiologically relevant, narrow state, enabling high RNA specificity. Moreover, the assembly of RNAs inside widens SmD1/D2/F/E/G, causes the release of Gemin2/SMN allosterically and allows SmD3/B to join. The assembly of SmD3/B further facilitates the release of Gemin2/SMN. This is the first to show negative cooperativity in snRNP assembly, which provides insights into RNA selection and the SMN complexs release. These findings reveal a basic mechanism of snRNP core assembly and facilitate pathogenesis studies of SMA.

molecular biology↗

bioRxiv2018

Cortical spike multiplexing using gamma frequency latencies

One of the fundamental problems in understanding the brain, in particular the cerebral cortex, is that we only have a partial understanding of the basic communication protocols that underlie signal transmission. This makes it difficult to interpret the significance of particular phenomena such as basic firing patterns and oscillations at different frequencies. There are, of course, useful models. Motivated by single-cell recording technology, Poisson statistics of cortical action potentials have long been a basic component in models of signal representation in the cortex. However, it is increasingly difficult to integrate Poisson spiking with spike timing signals in the gamma frequency spectrum. A potential way forward is being sparked by new technologies that allow the exploration of very low-level communication strategies. Specifically, the voltage potential of a cells soma now can be recorded with very high fidelity in vivo, allowing correlation of its fine structure to be correlated with behaviors. To interpret this data, we have developed a unified model (gamma spike multiplexing, or GSM) wherein a cells somatic gamma frequencies can modulate the generation of action potentials. Such spikes can be seen as the basis for a general-purpose method of modulating fast communication in cortical networks. In particular, the model has several important advantages over traditional formalisms: 1) It allows multiple, independent processes to run in parallel, greatly increasing the processing capability of the cortex 2) Its processing speed is 102 to 103 times faster than population coding methods 3) Its processes are not bound to specific locations, but migrate across cortical cells as a function of time, facilitating the maintenance of cortical cell calibration.

neuroscience↗

bioRxiv2018

NDUFAB1 Protects Heart by Coordinating Mitochondrial Respiratory Complex and Supercomplex Assembly

The impairment of mitochondrial bioenergetics, often coupled with exaggerated reactive oxygen species (ROS) production, is emerging as a common mechanism in diseases of organs with a high demand for energy, such as the heart. Building a more robust cellular powerhouse holds promise for protecting these organs in stressful conditions. Here, we demonstrate that NDUFAB1 (NADH:ubiquinone oxidoreductase subunit AB1), acts as a powerful cardio-protector by enhancing mitochondrial energy biogenesis. In particular, NDUFAB1 coordinates the assembly of respiratory complexes I, II, and III and supercomplexes, conferring greater capacity and efficiency of mitochondrial energy metabolism. Cardiac-specific deletion of Ndufab1 in mice caused progressive dilated cardiomyopathy associated with defective bioenergetics and elevated ROS levels, leading to heart failure and sudden death. In contrast, transgenic overexpression of Ndufab1 effectively enhanced mitochondrial bioenergetics and protected the heart against ischemia-reperfusion injury. Our findings identify NDUFAB1 as a central endogenous regulator of mitochondrial energy and ROS metabolism and thus provide a potential therapeutic target for the treatment of heart failure and other mitochondrial bioenergetics-centered diseases.

cell biology↗

bioRxiv2018

Development of a Cx46 targeting strategy for cancer stem cells

Gap junction-mediated cell-cell communication enables tumor cells to synchronize the execution of complex processes. Despite the connexin family of gap junction proteins being considered tumor suppressors, we previously found that glioblastoma cancer stem cells (CSCs) express higher levels of Cx46 compared to non-stem tumor cells, and this was necessary and sufficient for CSC maintenance. To develop a Cx46 targeting strategy, we utilized point mutants to disrupt specific functions of Cx46 and found that gap junction coupling was the critical function of Cx46 for CSCs. Based on this finding, we screened a clinically relevant library of small molecules and identified clofazimine as an inhibitor of Cx46-specific cell-cell communication. Clofazimine attenuated proliferation, self-renewal, and tumor growth and synergized with temozolomide to induce apoptosis. These data suggest that combining clofazimine with standard-of-care therapies may target glioblastoma CSCs. Furthermore, these results demonstrate the importance of targeting cell-cell communication as an anti-cancer therapy.

cancer biology↗

bioRxiv2018

Disentangling locus of perceptual learning in the visual hierarchy of motion processing

Visual perceptual learning (VPL) can lead to long-lasting perceptual improvements. While the efficacy of VPL is well established, there is still a considerable debate about what mechanisms underlie the effects of VPL. Much of this debate concentrates on where along the visual processing hierarchy behaviorally relevant plasticity takes place. Here, we aimed to tackle this question in context of motion processing, a domain where links between behavior and processing hierarchy are well established. Specifically, we took advantage of an established transition from component-dependent representations at the earliest level to pattern-dependent representations at the middle-level of cortical motion processing. We trained two groups of participants on the same motion direction identification task using either grating or plaid stimuli. A set of pre- and post-training tests was used to determine the degree of learning specificity and generalizability. This approach allowed us to disentangle contributions from both low- and mid-level motion processing, as well as high-level cognitive changes. We observed a complete bi-directional transfer of learning between component and pattern stimuli as long as they shared the same apparent motion direction. This result indicates learning-induced plasticity at intermediate levels of motion processing. Moreover, we found that motion VPL is specific to the trained stimulus direction, speed, size, and contrast, highlighting the pivotal role of basic visual features in VPL, and diminishing the possibility of non-sensory decision-level enhancements. Taken together, our study psychophysically examined a variety of factors mediating motion VPL, and demonstrated that motion VPL most likely alters visual computation in the middle stage of motion processing.

neuroscience↗

bioRxiv2018

Flexible top-down modulation in human ventral temporal cortex

Visual neuroscientists have long characterized attention as inducing a scaling or additive effect on fixed parametric functions describing neural responses (e.g., contrast response functions). Here, we instead propose that top-down effects are more complex and manifest in ways that depend not only on attention but also other cognitive processes involved in executing a task. To substantiate this theory, we analyze fMRI responses in human ventral temporal cortex (VTC) in a study where stimulus eccentricity and cognitive task are varied. We find that as stimuli are presented farther into the periphery, bottom-up stimulus-driven responses decline but top-down attentional enhancement increases substantially. This disproportionate enhancement of weak responses cannot be easily explained by conventional models of attention. Furthermore, we find that attentional effects depend on the specific cognitive task performed by the subject, indicating the influence of additional cognitive processes other than attention (e.g., decision-making). The effects we observe replicate in an independent experiment from the same study, and also generalize to a separate study involving different stimulus manipulations (contrast and phase coherence). Our results suggest that a quantitative understanding of top-down modulation requires more nuanced and more precise characterization of multiple cognitive factors involved in completing a perceptual task.

neuroscience↗

bioRxiv2018

Structural Capacitance in Protein Evolution and Human Diseases

Canonical mechanisms of protein evolution include the duplication and diversification of pre-existing folds through genetic alterations that include point mutations, insertions, deletions, and copy number amplifications, as well as post-translational modifications that modify processes such as folding efficiency and cellular localization. Following a survey of the human mutation database, we have identified an additional mechanism, that we term structural capacitance, which results in the de novo generation of microstructure in previously disordered regions. We suggest that the potential for structural capacitance confers select proteins with the capacity to evolve over rapid timescales, facilitating saltatory evolution as opoposed to exclusively canonical Darwinian mechanisms. Our results implicate the elements of protein microstructure generated by this distinct mechanism in the pathogenesis of a wide variety of human diseases. The benefits of rapidly furnishing the potential for evolutionary change conferred by structural capacitance are consequently counterbalanced by this accompanying risk, with the extent of this determined by the host immune system. The phenomenon of structural capacitance has implications ranging from the ancestral diversification of protein folds to the engineering of synthetic proteins with enhanced evolvability.

biochemistry↗

bioRxiv2018

Predicting CTCF-mediated chromatin loops using CTCF-MP

The three dimensional organization of chromosomes within the cell nucleus is highly regulated. It is known that CTCF is an important architectural protein to mediate long-range chromatin loops. Recent studies have shown that the majority of CTCF binding motif pairs at chromatin loop anchor regions are in convergent orientation. However, it remains unknown whether the genomic context at the sequence level can determine if a convergent CTCF motif pair is able to form chromatin loop. In this paper, we directly ask whether and what sequence-based features (other than the motif itself) may be important to establish CTCF-mediated chromatin loops. We found that motif conservation measured by \"branch-of-origin\" that accounts for motif turn-over in evolution is an important feature. We developed a new machine learning algorithm called CTCF-MP based on word2vec to demonstrate that sequence-based features alone have the capability to predict if a pair of convergent CTCF motifs would form a loop. Together with functional genomic signals from CTCF ChIP-seq and DNase-seq, CTCF-MP is able to make highly accurate predictions on whether a convergent CTCF motif pair would form a loop in a single cell type and also across different cell types. Our work represents an important step further to understand the sequence determinants that may guide the formation of complex chromatin architectures.

bioinformatics↗

bioRxiv2018

Rapid and dynamic alternative splicing impacts the Arabidopsis cold response transcriptome

BackgroundPlants have adapted to tolerate and survive constantly changing environmental conditions by re-programming gene expression. The scale of the contribution of alternative splicing (AS) to stress responses has been underestimated due to limitations in RNA-seq analysis programs and poor representation of AS transcripts in plant databases. Significantly, the dynamics of the AS response have not been investigated but this is now possible with accurate transcript quantification programs and AtRTD2, a new, comprehensive transcriptome for Arabidopsis.\n\nResultsUsing ultra-deep RNA-sequencing of a time-course of Arabidopsis thaliana plants exposed to cold treatment, we identified 8,949 genes with altered expression of which 2,442 showed significant differential alternative splicing (DAS) and 1,647 genes were regulated only at the level of AS (DAS-only). The high temporal resolution demonstrated the rapid induction of both transcription and AS resulting in coincident waves of differential expression (transcription) and differential alternative splicing in the first 6-9 hours of cold. The differentially expressed and DAS gene sets were largely non-overlapping, each comprising thousands of genes. The dynamic analysis of AS identified genes with rapid and sensitive AS within 3 h of transfer to the cold (early AS genes), which were enriched for splicing and transcription factors. A detailed investigation of the novel cold-response DAS-only gene, U2B\"-LIKE, suggested that it regulates AS and is required for tolerance to freezing.\n\nConclusionsOur data indicate that transcription and AS are the major regulators of transcriptome reprogramming that together govern the physiological and survival responses of plants to low temperature.

genomics↗

bioRxiv2018

Generation of a novel growth-enhanced and reduced environmental impact transgenic pig strain

In pig production, insufficient feed digestion causes excessive nutrients such as phosphorus and nitrogen, which are then released to the environment. To address the issue of environmental emissions, we have established transgenic pigs harboring a single-copy quad-cistronic transgene and simultaneously expressing three microbial enzymes, {beta}-glucanase, xylanase, and phytase in the salivary glands. All the transgenic enzymes were successfully expressed, and the digestion of non-starch polysaccharides (NSPs) and phytate in the feedstuff was enhanced. Fecal nitrogen and phosphate outputs were reduced by 23%-46%, and growth rate improved by 23.4% (gilts) and 24.4% (boars) when the pigs were fed on a corn and soybean-based diet and high-NSP diet. The transgenic pigs showed a 11.5%- 14.5% improvement in feed conversion rate compared to the age-matched wild-type littermates. These findings indicate that transgenic pigs are promising resources for improving feed efficiency and reducing nutrient emissions to the environment.

biochemistry↗

bioRxiv2017

Single Molecule Sequencing of Cell-free DNA from Maternal Plasma for Noninvasive Trisomy Detection

The demand of non-invasive prenatal testing for autosomal aneuploidy using cell-free fetal DNA (cffDNA) in maternal plasma is a highly sought-after diagnostic, with a rapidly growing market. Current approaches developed by next generation sequencing (NGS) need PCR amplifcation during sample preparation, which results in amplification bias in GC-rich areas of the human genome. With these approaches, the minimum fetal fraction in maternal plasma is 4% for the small differences in circulating cfDNA between trisomic and disomic pregnancies to be detectable. In this paper, we performed single molecule sequencing of cell-free DNA from maternal plasma for noninvasive trisomy 13, 18 and 21 detections using the GenoCare platform. We found that single molecule sequencing is sensitive enough to detect these chromosome abnormalities when the fetal DNA fraction is as low as 2%. Compared to the Hiseq2500 platform, no significant GC bias was observed. The improved sensitivity and unbiased GC readout make GenoCare a promising platform for autosomal aneuploidy detections, even in the very early stage of pregnancy.

genomics↗

bioRxiv2017

PrimerServer: a high-throughput primer design and specificity-checking platform

SummaryDesigning specific primers for multiple sites across the whole genome is still challenging, especially in species with complex genomes. Here we present PrimerServer, a high-throughput primer design and specificity-checking platform with both web and command-line interfaces. This platform efficiently integrates site selection, primer design, specificity checking and data presentation. In our case study, PrimerServer achieved high accuracy and a fast running speed for a large number of sites, suggesting its potential for molecular biology applications such as molecular breeding or medical testing.\n\nAvailability and ImplementationSource code for PrimerServer is available at https://github.com/billzt/PrimerServer. A demo server is freely accessible at https://primerserver.org, with all major browsers supported.\n\nContactzhangrui@caas.cn or guosandui@caas.cn

bioinformatics↗

bioRxiv2017

Cyanophages exhibit rhythmic infection patterns under light-dark cycles

Most living organisms exhibit diurnal rhythms as an adaptation to the daily light-dark (diel) cycle. However, diurnal rhythms have not been found in viruses. Here, we studied the diel infection patterns of bacteriophages infecting the unicellular cyanobacteria Prochlorococcus and Synechococcus, which are the most abundant photosynthetic organisms in the oceans. With lab cultures, we found that cyanophages used three infection strategies in the dark: no adsorption, adsorption but no replication, and replication. Interestingly, the former two exhibited rhythmic infection patterns under light-dark cycles. We further showed in the South China Sea and the Western Pacific Ocean that cyanophage abundances varied rhythmically, with a peak at night. Moreover, diel transcriptional rhythms of many cyanophage genes were found in the North Pacific Subtropical Gyre, which also peaked at night. Our results suggested that cyanophage infection of Prochlorococcus is synchronized to the light-dark cycle, which may result in a synchronized release of dissolved organic matter to the marine food web.

microbiology↗

bioRxiv2017

Resequencing the Escherichia coli genome by GenoCare single molecule sequencing platform

Next generation sequencing (NGS) has revolutionized life sciences research. Recently, a new class of third-generation sequencing platforms has arrived to meet increasing demands in the clinic, capable of directly measuring DNA and RNA sequences at the single-molecule level without amplification. Here, we use the new GenoCare single molecule sequencing platform from Direct Genomics to resequence the E. coli genome and show comparable performance to the Illumina MiSeq system. Our platform detects single-molecule fluorescence by total internal reflection microscopy, with sequencing-by-synthesis chemistry. With a consensus sequence of 99.71% nucleotide identity to that of the Illumina MiSeq systems, GenoCare was determined to be a reliable platform for single-molecule sequencing, with strong potential for clinical applications.

genomics↗

bioRxiv2017

An improved experimental model of cystic hydatid disease in liver resembling natural infection route with stable growing dynamics and immune reaction

Cystic echinococcosis is an endemic parasitic infection in Xinjiang, China and is causing serious economic burdens and public health concerns. An experimental murine model in vivo for hepatic cystic echinococcosis was established in C57B/6 mice by injection with human protoscolices via the portal vein of three different concentrations. Mice were followed up 10 months by ultrasound, gross anatomy, pathological and immunological examinations. The protoscolice migration in portal vein, hydatid cyst growth, host immune reaction and hepatic histopathology were examed periodicly. The infection rate of the mice in the high, medium, and low concentration groups were 90%, 100%, and 63.6%, respectively. The protoscolices migrate in the portal vein with blood flow, settle in the liver and develop into orthotopic hepatic hydatid cysts, resembling the natural infection route and course. This study established an improved experimental model of low biohazard risk but stable growing dynamics and immune reaction. It is especially useful for new anti-parasite medication trials agains hydatid disease.\n\nSummary statementAn experimental murine model of cystic echinococcosis was set up. This orthotopic model resembles primary infection route and natural infectious course with low biohazard risk and high efficiency.

pathology↗