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At least 307 records · Page 17

Binding profiles for 961 Drosophila and C. elegans transcription factors reveal tissue-specific regulatory relationships

A catalog of transcription factor (TF) binding sites in the genome is critical for deciphering regulatory relationships. Here, we present the culmination of the efforts of the modENCODE (model organism Encyclopedia of DNA Elements) and modERN (model organism Encyclopedia of Regulatory Networks) consortia to systematically assay TF binding events in vivo in two major model organisms,Drosophila melanogaster(fly) andCaenorhabditis elegans(worm). These data sets comprise 605 TFs identifying 3.6 M sites in the fly and 356 TFs identifying 0.9 M sites in the worm, and represent the majority of the regulatory space in each genome. We demonstrate that TFs associate with chromatin in clusters termed “metapeaks,” that larger metapeaks have characteristics of high-occupancy target (HOT) regions, and that the importance of consensus sequence motifs bound by TFs depends on metapeak size and complexity. Combining ChIP-seq data with single-cell RNA-seq data in a machine-learning model identifies TFs with a prominent role in promoting target gene expression in specific cell types, even differentiating between parent–daughter cells during embryogenesis. These data are a rich resource for the community that should fuel and guide future investigations into TF function. To facilitate data accessibility and utility, all strains expressing green fluorescent protein (GFP)-tagged TFs are available at the stock centers for each organism. The chromatin immunoprecipitation sequencing data are available through the ENCODE Data Coordinating Center, GEO, and through a direct interface that provides rapid access to processed data sets and summary analyses, as well as widgets to probe the cell-type-specific TF–target relationships.

Biochemistry & Molecular Biology↗

Evaluation of Potential In Vitro Recombination Events in Codon Deoptimized FMDV Strains

Codon deoptimization (CD) has been recently used as a possible strategy to derive foot-and-mouth disease (FMD) live-attenuated vaccine (LAV) candidates containing DIVA markers. However, reversion to virulence, or loss of DIVA, from possible recombination with wild-type (WT) strains has yet to be analyzed. An in vitro assay was developed to quantitate the levels of recombination between WT and a prospective A24-P2P3 partially deoptimized LAV candidate. By using two genetically engineered non-infectious RNA templates, we demonstrate that recombination can occur within non-deoptimized viral genomic regions (i.e., 3'end of P3 region). The sequencing of single plaque recombinants revealed a variety of genome compositions, including full-length WT sequences at the consensus level and deoptimized sequences at the sub-consensus/consensus level within the 3'end of the P3 region. Notably, after further passage, two recombinants that contained deoptimized sequences evolved to WT. Overall, recombinants featuring large stretches of CD or DIVA markers were less fit than WT viruses. Our results indicate that the developed assay is a powerful tool to evaluate the recombination of FMDV genomes in vitro and should contribute to the improved design of FMDV codon deoptimized LAV candidates.

59 BASIC BIOLOGICAL SCIENCES↗

Comparative genomics of pyrophilous fungi reveals a link between fire events and developmental genes

Forest fires generate a large amount of carbon that remains resident on the site as dead and partially ‘pyrolysed’ (i.e. burnt) material that has long residency times and constitutes a significant pool in fire–prone ecosystems. In addition, fire–induced hydrophobic soil layers, caused by condensation of pyrolysed waxes and lipids, increase post–fire erosion and can lead to long–term productivity losses. A small set of pyrophilous fungi dominate post–fire soils and are likely to be involved with the degradation of all these compounds, yet almost nothing is currently known about what these fungi do or the metabolic processes they employ. In this study, we sequenced and analyzed genomes from fungi isolated after Rim fire near Yosemite National Park in 2013 and showed the enrichment/expansion of CAZymes and families known to be involved in fruiting body initiation when compared to other basidiomycete fungi. We found gene families potentially involved in the degradation of the hydrophobic layer and pyrolysed organic matter, such as hydrophobic surface binding proteins, laccases (AA1_1), xylanases (GH10, GH11), fatty acid desaturases and tannases. Furthermore, pyrophilous fungi are important actors to restate the soil's functional capabilities.

59 BASIC BIOLOGICAL SCIENCES↗

Radioimaging for real-time tracking of high-voltage breakdown

Development of a radioimaging diagnostic for high-voltage component reliability testing and electrical breakdown computational model validation is described. Radioimaging has its roots in radio astronomy, where aperture synthesis (also known as synthesis imaging) has been utilized for decades to image radio sources far from Earth. Radioimaging as described herein, in contrast, seeks to image radio sources in close proximity to its receivers (i.e., in a laboratory environment). Here it is shown that corona discharge, a non-destructive precursor to catastrophic (thermal) arc discharge, electromagnetically radiates strongly within a 250 kHz – 2.5 GHz bandwidth, and is readily detected and located by postprocessing the received radio signals. The ability of radioimaging to detect both corona and arc discharge (grouped together herein as high voltage breakdown or HVB) makes it a valuable tool for 100% HVB detection in materials, components, and devices, and has the ability to indicate electrical weakness (via corona detection) prior to a destructive arc discharge event. Radioimaging enables HVB to be located both internal and external to dielectric components under test in near-real-time, with multiple and/or extended HVB events located simultaneously. In contrast, existing non-destructive diagnostics (at the time of this writing) either indicate electrical breakdown without resolving failure locations (e.g., current, voltage, and chemical measurements), locate external HVB (e.g., high-speed optical and ultraviolet (UV) measurements or photography), or locate both external and internal HVB but with low fidelity (e.g., a single HVB source can be located by existing time-of-arrival (TOA) UHF or acoustic emissions). Radioimaging instead creates a sequence of high-fidelity images similar to an optical high-speed camera but at radiofrequencies (RF), and is not limited to two-dimensions. Moreover, radioimaging has already served one internal and two external industry customers, the results of which are detailed in this report. The radioimaging results described herein were part of a three-year effort funded by the Sandia Lab Directed Research and Development (LDRD) program within the Radiation, Electromagnetic, High Energy Density Science (REHEDS) investment area.

47 OTHER INSTRUMENTATION↗

Convergent evolution and horizontal gene transfer in Arctic Ocean microalgae

Microbial communities in the world ocean are affected strongly by oceanic circulation, creating characteristic marine biomes. The high connectivity of most of the ocean makes it difficult to disentangle selective retention of colonizing genotypes (with traits suited to biome specific conditions) from evolutionary selection, which would act on founder genotypes over time. The Arctic Ocean is exceptional with limited exchange with other oceans and ice covered since the last ice age. To test whether Arctic microalgal lineages evolved apart from algae in the global ocean, we sequenced four lineages of microalgae isolated from Arctic waters and sea ice. Here we show convergent evolution and highlight geographically limited HGT as an ecological adaptive force in the form of PFAM complements and horizontal acquisition of key adaptive genes. Notably, ice-binding proteins were acquired and horizontally transferred among Arctic strains. A comparison with Tara Oceans metagenomes and metatranscriptomes confirmed mostly Arctic distributions of these IBPs. The phylogeny of Arctic-specific genes indicated that these events were independent of bacterial-sourced HGTs in Antarctic Southern Ocean microalgae.

59 BASIC BIOLOGICAL SCIENCES↗

The EBLM project – VIII. First results for M-dwarf mass, radius, and effective temperature measurements using CHEOPS light curves

ABSTRACT The accuracy of theoretical mass, radius, and effective temperature values for M-dwarf stars is an active topic of debate. Differences between observed and theoretical values have raised the possibility that current theoretical stellar structure and evolution models are inaccurate towards the low-mass end of the main sequence. To explore this issue, we use the CHEOPS satellite to obtain high-precision light curves of eclipsing binaries with low-mass stellar companions. We use these light curves combined with the spectroscopic orbit for the solar-type companion to measure the mass, radius, and effective temperature of the M-dwarf star. Here, we present the analysis of three eclipsing binaries. We use the pycheops data analysis software to fit the observed transit and eclipse events of each system. Two of our systems were also observed by the TESS satellite – we similarly analyse these light curves for comparison. We find consistent results between CHEOPS and TESS, presenting three stellar radii and two stellar effective temperature values of low-mass stellar objects. These initial results from our on-going observing programme with CHEOPS show that we can expect to have ∼24 new mass, radius, and effective temperature measurements for very low-mass stars within the next few years.

Swayne, M. I.↗

Cytochrome b5 diversity in green lineages preceded the evolution of syringyl lignin biosynthesis

Abstract Lignin production marked a milestone in vascular plant evolution, and the emergence of syringyl (S) lignin is lineage specific. S-lignin biosynthesis in angiosperms, mediated by ferulate 5-hydroxylase (F5H, CYP84A1), has been considered a recent evolutionary event. F5H uniquely requires the cytochrome b5 protein CB5D as an obligatory redox partner for catalysis. However, it remains unclear how CB5D functionality originated and whether it coevolved with F5H. We reveal here the ancient evolution of CB5D-type function supporting F5H-catalyzed S-lignin biosynthesis. CB5D emerged in charophyte algae, the closest relatives of land plants, and is conserved and proliferated in embryophytes, especially in angiosperms, suggesting functional diversification of the CB5 family before terrestrialization. A sequence motif containing acidic amino residues in Helix 5 of the CB5 heme-binding domain contributes to the retention of CB5D function in land plants but not in algae. Notably, CB5s in the S-lignin–producing lycophyte Selaginella lack these residues, resulting in no CB5D-type function. An independently evolved S-lignin biosynthetic F5H (CYP788A1) in Selaginella relies on NADPH-dependent cytochrome P450 reductase as sole redox partner, distinct from angiosperms. These results suggest that angiosperm F5Hs coopted the ancient CB5D, forming a modern cytochrome P450 monooxygenase system for aromatic ring meta-hydroxylation, enabling the reemergence of S-lignin biosynthesis in angiosperms.

59 BASIC BIOLOGICAL SCIENCES↗

Machine Learning of All Mycobacterium tuberculosis H37Rv RNA-seq Data Reveals a Structured Interplay between Metabolism, Stress Response, and Infection

Mycobacterium tuberculosis is one of the most consequential human bacterial pathogens, posing a serious challenge to 21st century medicine. A key feature of its pathogenicity is its ability to adapt its transcriptional response to environmental stresses through its transcriptional regulatory network (TRN). While many studies have sought to characterize specific portions of the M. tuberculosis TRN, and some studies have performed system-level analysis, few have been able to provide a network-based model of the TRN that also provides the relative shifts in transcriptional regulator activity triggered by changing environments. Here, we compiled a compendium of nearly 650 publicly available, high quality M. tuberculosis RNA-sequencing data sets and applied an unsupervised machine learning method to obtain a quantitative, top-down TRN. It consists of 80 independently modulated gene sets known as “iModulons,” 41 of which correspond to known regulons. These iModulons explain 61% of the variance in the organism’s transcriptional response. We show that iModulons (i) reveal the function of poorly characterized regulons, (ii) describe the transcriptional shifts that occur during environmental changes such as shifting carbon sources, oxidative stress, and infection events, and (iii) identify intrinsic clusters of regulons that link several important metabolic systems, including lipid, cholesterol, and sulfur metabolism. This transcriptome-wide analysis of the M. tuberculosis TRN informs future research on effective ways to study and manipulate its transcriptional regulation and presents a knowledge-enhanced database of all published high-quality RNA-seq data for this organism to date.

59 BASIC BIOLOGICAL SCIENCES↗

Use of Fluorescent Protein Reporters for Assessing and Detecting Genome Editing Reagents and Transgene Expression in Plants

Fluorescent protein reporters have been widely used for monitoring the expression of target genes in various engineered organisms. Although a wide range of analytical approaches (e.g., genotyping PCR, digital PCR, DNA sequencing) have been utilized to detect and identify genome editing reagents and transgene expression in genetically modified plants, these methods are usually limited to use in the late stages of plant transformation and can only be used invasively. Here we describe GFP- and eYGFPuv-based strategies and methods for assessing and detecting genome editing reagents and transgene expression in plants, including protoplast transformation, leaf infiltration, and stable transformation. These methods and strategies enable easy, noninvasive screening of genome editing and transgenic events in plants.

Yuan, Guoliang↗

Spatiotemporal forecasting of the edge localized modes in tokamak plasmas using neural networks

Artificial intelligence techniques have been increasingly adopted by the plasma and fusion science to address problems like plasma reconstruction, surrogate modeling, and tokamak/stellarator optimization. A key focus in sustained fusion research is the prediction and mitigation of edge-localized-modes (ELMs), instabilities that occur in short, periodic bursts and can cause erosion to the tokamak vessel wall. Recent research has demonstrated the power of neural networks in approximating continuous functions. In this work, we build spatiotemporal forecasting models that can predict the onset of ELMs and their evolution at early stages. We leverage recent advances in generative modeling, sequence-to-sequence modeling, and Fourier neural operators to propose architectures and training strategies that can learn to forecast short to long term dynamics of the noisy signals due to ELMs. We benchmark the developed model against a state-of-the-art foundation model using the beam emission spectroscopy (BES) data that captures the plasma fluctuations due to ELMs over a 8 x 8 spatial grid. Our models demonstrate high accuracy, outperforming the baselines, in predicting the evolution of BES signals during ELM events. Furthermore, the developed models exhibit high accuracy in predicting the rapid rise and relaxation of the signals due to ELMs within 30–80 µs.

edge localized modes↗

Sandwich Hybridization Assay for In Situ Real-Time Cyanobacterial Detection and Monitoring: A Review

As cyanobacterial harmful algal bloom (cHAB) events increase in scale, severity, frequency, and duration around the world, rapid and accurate monitoring and characterization tools have become critically essential for regulatory and management decision-making. The composition of cHAB-forming cyanobacteria community can change significantly over time and space and be altered by sample preservation and transportation, making in situ monitoring necessary to obtain real-time and localized information. Sandwich hybridization assay (SHA) utilizes capture oligonucleotide probes for sensitive detection of target-specific nucleic acid sequences. As an amplification-free molecular biology technology, SHA can be adapted for in-situ, real-time or near real-time detection and qualitatively or semi-quantitatively monitoring of cHAB-forming cyanobacteria, owing to its characteristics such as being rapid, portable, inexpensive, and amenable to automation, high sensitivity, specificity and robustness, and multiplexing (i.e., detecting multiple targets simultaneously). Despite its successful application in the monitoring of marine and freshwater phytoplankton, there is still room for improvement. The ability to identify a cHAB community rapidly would decrease delays in cyanotoxin analyses, reduce costs, and increase sample throughput, allowing for timely actions to improve environmental and human health and the understanding of short- and long-term bloom dynamics. Real-time detection and quantitation of HAB-forming cyanobacteria is essential for improving environmental and public health and reducing associated costs. We review and propose to apply SHA for in situ cHABs monitoring.

59 BASIC BIOLOGICAL SCIENCES↗

A study of convective core overshooting as a function of stellar mass based on two-dimensional hydrodynamical simulations

ABSTRACT We perform two-dimensional (2D) numerical simulations of core convection for zero-age main-sequence stars covering a mass range from 3 to 20 M⊙. The simulations are performed with the fully compressible time-implicit code music. We study the efficiency of overshooting, which describes the ballistic process of convective flows crossing a convective boundary, as a function of stellar mass and luminosity. We also study the impact of artificially increasing the stellar luminosity for 3 M⊙ models. The simulations cover hundreds to thousands of convective turnover time-scales. Applying the framework of extreme plume events previously developed for convective envelopes, we derive overshooting lengths as a function of stellar masses. We find that the overshooting distance (dov) scales with the stellar luminosity (L) and the convective core radius (rconv). We derive a scaling law $d_{\rm ov} \propto L^{1/3} r_{\rm conv}^{1/2}$, which is implemented in a one-dimensional stellar evolution code and the resulting stellar models are compared to observations. The scaling predicts values for the overshooting distance that significantly increase with stellar mass, in qualitative agreement with observations. Quantitatively, however, the predicted values are underestimated for masses ≳10 M⊙. Our 2D simulations show the formation of a nearly adiabatic layer just above the Schwarzschild boundary of the convective core, as exhibited in recent three-dimensional simulations of convection. The most luminous models show a growth in size with time of the nearly adiabatic layer. This growth seems to slow down as the upper edge of the nearly adiabatic layer gets closer to the maximum overshooting length and as the simulation time exceeds the typical thermal diffusive time-scale in the overshooting layer.

79 ASTRONOMY AND ASTROPHYSICS↗

An Asymmetric Eclipse Seen toward the Pre-main-sequence Binary System V928 Tau

K2 observations of the weak-lined T Tauri binary V928 Tau A and B show the detection of a single, asymmetric eclipse, which may be due to a previously unknown substellar companion eclipsing one component of the binary with an orbital period >66 days. Over an interval of about 9 hr, one component of the binary dims by around 60%, returning to its normal brightness about 5 hr later. From modeling of the eclipse shape, we find evidence that the eclipsing companion may be surrounded by a disk or a vast ring system. The modeled disk has a radius of 0.9923 ± 0.0005 R∗, with an inclination of 56.°78 ± 0.°03, a tilt of 41.°22 ± 0.°05, an impact parameter of -0.2506 ± 0.0002 R∗, and an opacity of 1.00. The occulting disk must also move at a transverse velocity of 6.637 ± 0.002 R∗ day -1 , which, depending on whether it orbits V928 Tau A or B, corresponds to approximately 73.53 or 69.26 km s -1 . A search in ground-based archival data reveals additional dimming events, some of which suggest periodicity, but no unambiguous period associated with the eclipse observed by K2. We present a new epoch of astrometry that is used to further refine the orbit of the binary, presenting a new lower bound of 67 yr, and constraints on the possible orbital periods of the eclipsing companion. The binary is also separated by 18' (~2250 au) from the lower-mass CFHT-BD-Tau 7, which is likely associated with V928 Tau A and B. We also present new high-dispersion optical spectroscopy that we use to characterize the unresolved stellar binary.

79 ASTRONOMY AND ASTROPHYSICS↗

An End-To-End Earthquake Detection Method for Joint Phase Picking and Association Using Deep Learning

Earthquake monitoring by seismic networks typically involves a workflow consisting of phase detection/picking, association, and location tasks. In recent years, the accuracy of these individual stages has been improved through the use of machine learning techniques. Here, in this study, we introduce a new end-to-end approach that improves overall earthquake detection accuracy by jointly optimizing each stage of the detection pipeline. We propose a neural network architecture for the task of multi-station processing of seismic waveforms recorded over a seismic network. This end-to-end architecture consists of three sub-networks: a backbone network that extracts features from raw waveforms, a phase picking sub-network that picks P- and S-wave arrivals based on these features, and an event detection sub-network that aggregates the features from multiple stations to associate and detect earthquakes across a seismic network. We use these sub-networks together with a shift-and-stack module based on back-projection that introduces kinematic constraints on arrival times, allowing the neural network model to generalize to different velocity models and to variable station geometry in seismic networks. We evaluate our proposed method on the STanford EArthquake Dataset (STEAD) and on the 2019 Ridgecrest, CA earthquake sequence. The results demonstrate that our end-to-end approach can effectively pick P- and S-wave arrivals and achieve earthquake detection accuracy rivaling that of other state-of-the-art approaches. Because our approach preserves information across tasks in the detection pipeline, it has the potential to outperform approaches that do not.

58 GEOSCIENCES↗

Nuclear Data Sheets for A=48

Experimental nuclear structure data from various reactions and decays are compiled and evaluated for all known nuclides with mass number A=48 (S, Cl, Ar, K, Ca, Sc, Ti, V, Cr, Mn, Fe, Co, Ni). For each nuclide, detailed evaluated nuclear structure information is presented for each individual reaction and decay, and the best values combining all available data are recommended for spectroscopic properties, such as level energies, half-lives, γ-ray energies and intensities, decay radiations. No excited states have been identified in 48 S, 48 Cl, 48 Co and 48 Ni, among which the first three even have no measured ground-state half-lives and decay modes. For 48 Fe, a level scheme with a sequence of excited states was established for the first time from a recent one-neutron removal measurement by 2021Ya33. Data for excited states in 48 Ar, 48 K and 48 Mn remain limited; no decay scheme has been measured for 48 Ar and 48 K yet, while the decay scheme for 48 Ni is incomplete due to unobserved levels. 48 Ti is the most studied nuclide through various reactions and decays, followed by 48 V, 40 Ca, 48 Sc, and 48 Cr, among which no decay event to 48 Sc has been observed and the decay schemes for 48 V and 48 Cr are considered as incomplete. Furthermore, this work supersedes the previous full evaluations of A=48: 2006Bu08, 1993Bu04, 1985Al14, 1978Be01.

73 NUCLEAR PHYSICS AND RADIATION PHYSICS↗

Predicting the chemical space of fungal polyketides by phylogeny-based bioinformatics analysis of polyketide synthase-nonribosomal peptide synthetase and its modification enzymes

Fungal polyketide synthase (PKS)–nonribosomal peptide synthetase (NRPS) hybrids are key enzymes for synthesizing structurally diverse hybrid natural products (NPs) with characteristic biological activities. Predicting their chemical space is of particular importance in the field of natural product chemistry. However, the unexplored programming rule of the PKS module has prevented prediction of its chemical structure based on amino acid sequences. Here, we conducted a phylogenetic analysis of 884 PKS–NRPS hybrids and a modification enzyme analysis of the corresponding biosynthetic gene cluster, revealing a hidden relationship between its genealogy and core structures. This unexpected result allowed us to predict 18 biosynthetic gene cluster (BGC) groups producing known carbon skeletons (number of BGCs; 489) and 11 uncharacterized BGC groups (171). The limited number of carbon skeletons suggests that fungi tend to select PK skeletons for survival during their evolution. The possible involvement of a horizontal gene transfer event leading to the diverse distribution of PKS–NRPS genes among fungal species is also proposed. This study provides insight into the chemical space of fungal PKs and the distribution of their biosynthetic gene clusters.

37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CH↗

Identification of genetic interactions with priB links the PriA/PriB DNA replication restart pathway to double-strand DNA break repair in Escherichia coli

Collisions between DNA replication complexes (replisomes) and impediments such as damaged DNA or proteins tightly bound to the chromosome lead to premature dissociation of replisomes at least once per cell cycle in Escherichia coli. Left unrepaired, these events produce incompletely replicated chromosomes that cannot be properly partitioned into daughter cells. DNA replication restart, the process that reloads replisomes at prematurely terminated sites, is therefore essential in E. coli and other bacteria. Three replication restart pathways have been identified in E. coli: PriA/PriB, PriA/PriC, and PriC/Rep. A limited number of genetic interactions between replication restart and other genome maintenance pathways have been defined, but a systematic study placing replication restart reactions in a broader cellular context has not been performed. We have utilized transposon-insertion sequencing to identify new genetic interactions between DNA replication restart pathways and other cellular systems. Known genetic interactors with the priB replication restart gene (uniquely involved in the PriA/PriB pathway) were confirmed and several novel priB interactions were discovered. Targeted genetic and imaging-based experiments with priB and its genetic partners revealed significant double-strand DNA break accumulation in strains with mutations in dam, rep, rdgC, lexA, or polA. Modulating the activity of the RecA recombinase partially suppressed the detrimental effects of rdgC or lexA mutations in ΔpriB cells. Taken together, our results highlight roles for several genes in double-strand DNA break homeostasis and define a genetic network that facilitates DNA repair/processing upstream of PriA/PriB-mediated DNA replication restart in E. coli.

59 BASIC BIOLOGICAL SCIENCES↗

Global metagenomics reveals plastid diversity and unexplored algal lineages

Photosynthetic organelles in eukaryotes originated through primary endosymbiosis with a cyanobacterium, an event that profoundly shaped the evolutionary landscape of the eukaryotic tree of life. Primary plastids in Archaeplastida, especially in cultivable plants and algae, contribute most to known plastid diversity. Secondary and higher-order endosymbiosis, involving eukaryotic hosts and algal endosymbionts, further spread photosynthesis among protists within the CASH lineages (Cryptophyta, Alveolata, Stramenopila, and Haptophyta). Despite various hypotheses explaining secondary plastid evolution and distribution, empirical support remains limited. Here, we employ cultivation-independent global metagenomics to expand plastid diversity and investigate plastid origins. We capture 1,027 plastid sequences, including 300 novel sequences belonging to previously unsequenced plastids and representing yet-to-be described microeukaryotes. This includes a new lineage that offers insights into plastid evolution in haptophytes and cryptophytes. Our results confirm that Archaeplastida plastids originate from an early branching cyanobacterial lineage closely related to Gloeomargaritales and identify the closest extant relative of Paulinella plastids. Additionally, our findings suggest two independent origins of secondary red-algal plastids, contributing to plastid diversity in CASH lineages and challenging the prevailing model of single secondary plastid origin. Our study highlights the importance of metagenomic data in uncovering biological diversity and advancing understanding of plastid relationships across photosynthetic eukaryotes.

59 BASIC BIOLOGICAL SCIENCES↗