BioSciences Publications

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BioSciences faculty publications. For works published before Summer 2014, please see the Biochemistry & Cell Biology and Ecology & Evolutionary Biology collections.

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    A deep learning solution for crystallographic structure determination
    (International Union of Crystallography, 2023) Pan, T.; Jin, S.; Miller, M. D.; Kyrillidis, A.; Phillips, G. N.
    The general de novo solution of the crystallographic phase problem is difficult and only possible under certain conditions. This paper develops an initial pathway to a deep learning neural network approach for the phase problem in protein crystallography, based on a synthetic dataset of small fragments derived from a large well curated subset of solved structures in the Protein Data Bank (PDB). In particular, electron-density estimates of simple artificial systems are produced directly from corresponding Patterson maps using a convolutional neural network architecture as a proof of concept.
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    Endogenous viral elements reveal associations between a non-retroviral RNA virus and symbiotic dinoflagellate genomes
    (Springer Nature, 2023) Veglia, Alex J.; Bistolas, Kalia S. I.; Voolstra, Christian R.; Hume, Benjamin C. C.; Ruscheweyh, Hans-Joachim; Planes, Serge; Allemand, Denis; Boissin, Emilie; Wincker, Patrick; Poulain, Julie; Moulin, Clémentine; Bourdin, Guillaume; Iwankow, Guillaume; Romac, Sarah; Agostini, Sylvain; Banaigs, Bernard; Boss, Emmanuel; Bowler, Chris; de Vargas, Colomban; Douville, Eric; Flores, Michel; Forcioli, Didier; Furla, Paola; Galand, Pierre E.; Gilson, Eric; Lombard, Fabien; Pesant, Stéphane; Reynaud, Stéphanie; Sunagawa, Shinichi; Thomas, Olivier P.; Troublé, Romain; Zoccola, Didier; Correa, Adrienne M. S.; Vega Thurber, Rebecca L.
    Endogenous viral elements (EVEs) offer insight into the evolutionary histories and hosts of contemporary viruses. This study leveraged DNA metagenomics and genomics to detect and infer the host of a non-retroviral dinoflagellate-infecting +ssRNA virus (dinoRNAV) common in coral reefs. As part of the Tara Pacific Expedition, this study surveyed 269 newly sequenced cnidarians and their resident symbiotic dinoflagellates (Symbiodiniaceae), associated metabarcodes, and publicly available metagenomes, revealing 178 dinoRNAV EVEs, predominantly among hydrocoral-dinoflagellate metagenomes. Putative associations between Symbiodiniaceae and dinoRNAV EVEs were corroborated by the characterization of dinoRNAV-like sequences in 17 of 18 scaffold-scale and one chromosome-scale dinoflagellate genome assembly, flanked by characteristically cellular sequences and in proximity to retroelements, suggesting potential mechanisms of integration. EVEs were not detected in dinoflagellate-free (aposymbiotic) cnidarian genome assemblies, including stony corals, hydrocorals, jellyfish, or seawater. The pervasive nature of dinoRNAV EVEs within dinoflagellate genomes (especially Symbiodinium), as well as their inconsistent within-genome distribution and fragmented nature, suggest ancestral or recurrent integration of this virus with variable conservation. Broadly, these findings illustrate how +ssRNA viruses may obscure their genomes as members of nested symbioses, with implications for host evolution, exaptation, and immunity in the context of reef health and disease.
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    Stony coral tissue loss disease induces transcriptional signatures of in situ degradation of dysfunctional Symbiodiniaceae
    (Springer Nature, 2023) Beavers, Kelsey M.; Van Buren, Emily W.; Rossin, Ashley M.; Emery, Madison A.; Veglia, Alex J.; Karrick, Carly E.; MacKnight, Nicholas J.; Dimos, Bradford A.; Meiling, Sonora S.; Smith, Tyler B.; Apprill, Amy; Muller, Erinn M.; Holstein, Daniel M.; Correa, Adrienne M. S.; Brandt, Marilyn E.; Mydlarz, Laura D.
    Stony coral tissue loss disease (SCTLD), one of the most pervasive and virulent coral diseases on record, affects over 22 species of reef-building coral and is decimating reefs throughout the Caribbean. To understand how different coral species and their algal symbionts (family Symbiodiniaceae) respond to this disease, we examine the gene expression profiles of colonies of five species of coral from a SCTLD transmission experiment. The included species vary in their purported susceptibilities to SCTLD, and we use this to inform gene expression analyses of both the coral animal and their Symbiodiniaceae. We identify orthologous coral genes exhibiting lineage-specific differences in expression that correlate to disease susceptibility, as well as genes that are differentially expressed in all coral species in response to SCTLD infection. We find that SCTLD infection induces increased expression of rab7, an established marker of in situ degradation of dysfunctional Symbiodiniaceae, in all coral species accompanied by genus-level shifts in Symbiodiniaceae photosystem and metabolism gene expression. Overall, our results indicate that SCTLD infection induces symbiophagy across coral species and that the severity of disease is influenced by Symbiodiniaceae identity.
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    How to tuna fish: constraint, convergence, and integration in the neurocranium of pelagiarian fishes
    (Oxford University Press, 2023) Knapp, Andrew; Rangel-de Lázaro, Gizéh; Friedman, Matt; Johanson, Zerina; Evans, Kory M; Giles, Sam; Beckett, Hermione T; Goswami, Anjali
    Morphological evolution of the vertebrate skull has been explored across a wide range of tetrapod clades using geometric morphometrics, but the application of these methods to teleost fishes, accounting for roughly half of all vertebrate species, has been limited. Here we present the results of a study investigating 3D morphological evolution of the neurocranium across 114 species of Pelagiaria, a diverse clade of open-ocean teleost fishes that includes tuna and mackerel. Despite showing high shape disparity overall, taxa from all families fall into three distinct morphological clusters. Convergence in shape within clusters is high, and phylogenetic signal in shape data is significant but low. Neurocranium shape is significantly correlated with body elongation and significantly but weakly correlated with size. Diet and habitat depth are weakly correlated with shape, and nonsignificant after accounting for phylogeny. Evolutionary integration in the neurocranium is high, suggesting that convergence in skull shape and the evolution of extreme morphologies are associated with the correlated evolution of neurocranial elements. These results suggest that shape evolution in the pelagiarian neurocranium reflects the extremes in elongation found in body shape but is constrained along relatively few axes of variation, resulting in repeated evolution toward a restricted range of morphologies.
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    Building consensus around the assessment and interpretation of Symbiodiniaceae diversity
    (PeerJ, Inc, 2023) Davies, Sarah W.; Gamache, Matthew H.; Howe-Kerr, Lauren I.; Kriefall, Nicola G.; Baker, Andrew C.; Banaszak, Anastazia T.; Bay, Line Kolind; Bellantuono, Anthony J.; Bhattacharya, Debashish; Chan, Cheong Xin; Claar, Danielle C.; Coffroth, Mary Alice; Cunning, Ross; Davy, Simon K.; Campo, Javier del; Díaz-Almeyda, Erika M.; Frommlet, Jörg C.; Fuess, Lauren E.; González-Pech, Raúl A.; Goulet, Tamar L.; Hoadley, Kenneth D.; Howells, Emily J.; Hume, Benjamin C. C.; Kemp, Dustin W.; Kenkel, Carly D.; Kitchen, Sheila A.; LaJeunesse, Todd C.; Lin, Senjie; McIlroy, Shelby E.; McMinds, Ryan; Nitschke, Matthew R.; Oakley, Clinton A.; Peixoto, Raquel S.; Prada, Carlos; Putnam, Hollie M.; Quigley, Kate; Reich, Hannah G.; Reimer, James Davis; Rodriguez-Lanetty, Mauricio; Rosales, Stephanie M.; Saad, Osama S.; Sampayo, Eugenia M.; Santos, Scott R.; Shoguchi, Eiichi; Smith, Edward G.; Stat, Michael; Stephens, Timothy G.; Strader, Marie E.; Suggett, David J.; Swain, Timothy D.; Tran, Cawa; Traylor-Knowles, Nikki; Voolstra, Christian R.; Warner, Mark E.; Weis, Virginia M.; Wright, Rachel M.; Xiang, Tingting; Yamashita, Hiroshi; Ziegler, Maren; Correa, Adrienne M. S.; Parkinson, John Everett
    Within microeukaryotes, genetic variation and functional variation sometimes accumulate more quickly than morphological differences. To understand the evolutionary history and ecology of such lineages, it is key to examine diversity at multiple levels of organization. In the dinoflagellate family Symbiodiniaceae, which can form endosymbioses with cnidarians (e.g., corals, octocorals, sea anemones, jellyfish), other marine invertebrates (e.g., sponges, molluscs, flatworms), and protists (e.g., foraminifera), molecular data have been used extensively over the past three decades to describe phenotypes and to make evolutionary and ecological inferences. Despite advances in Symbiodiniaceae genomics, a lack of consensus among researchers with respect to interpreting genetic data has slowed progress in the field and acted as a barrier to reconciling observations. Here, we identify key challenges regarding the assessment and interpretation of Symbiodiniaceae genetic diversity across three levels: species, populations, and communities. We summarize areas of agreement and highlight techniques and approaches that are broadly accepted. In areas where debate remains, we identify unresolved issues and discuss technologies and approaches that can help to fill knowledge gaps related to genetic and phenotypic diversity. We also discuss ways to stimulate progress, in particular by fostering a more inclusive and collaborative research community. We hope that this perspective will inspire and accelerate coral reef science by serving as a resource to those designing experiments, publishing research, and applying for funding related to Symbiodiniaceae and their symbiotic partnerships.
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    SARS-CoV-2 Exposure in Norway Rats (Rattus norvegicus) from New York City
    (American Society for Microbiology, 2023) Wang, Yang; Lenoch, Julianna; Kohler, Dennis; DeLiberto, Thomas J.; Tang, Cynthia Y.; Li, Tao; Tao, Yizhi Jane; Guan, Minhui; Compton, Susan; Zeiss, Caroline; Hang, Jun; Wan, Xiu-Feng
    Millions of Norway rats (Rattus norvegicus) inhabit New York City (NYC), presenting the potential for transmission of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) from humans to rats. We evaluated SARS-CoV-2 exposure among 79 rats captured from NYC during the fall of 2021. Our results showed that 13 of the 79 rats (16.5%) tested IgG- or IgM-positive, and partial SARS-CoV-2 genomes were recovered from all 4 rats that were qRT-PCR (reverse transcription-quantitative PCR)-positive. Genomic analyses suggest these viruses were associated with genetic lineage B, which was predominant in NYC in the spring of 2020 during the early pandemic period. To further investigate rat susceptibility to SARS-CoV-2 variants, we conducted a virus challenge study and showed that Alpha, Delta, and Omicron variants can cause infections in wild-type Sprague Dawley (SD) rats, including high replication levels in the upper and lower respiratory tracts and induction of both innate and adaptive immune responses. Additionally, the Delta variant resulted in the highest infectivity. In summary, our results indicate that rats are susceptible to infection with Alpha, Delta, and Omicron variants, and wild Norway rats in the NYC municipal sewer systems have been exposed to SARS-CoV-2. Our findings highlight the need for further monitoring of SARS-CoV-2 in urban rat populations and for evaluating the potential risk of secondary zoonotic transmission from these rat populations back to humans. IMPORTANCE The host tropism expansion of SARS-CoV-2 raises concern for the potential risk of reverse-zoonotic transmission of emerging variants into rodent species, including wild rat species. In this study, we present both genetic and serological evidence for SARS-CoV-2 exposure to the New York City wild rat population, and these viruses may be linked to the viruses that were circulating during the early stages of the pandemic. We also demonstrated that rats are susceptible to additional variants (i.e., Alpha, Delta, and Omicron) that have been predominant in humans and that susceptibility to infection varies by variant. Our findings highlight the reverse zoonosis of SARS-CoV-2 to urban rats and the need for further monitoring of SARS-CoV-2 in rat populations for potential secondary zoonotic transmission to humans.
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    Examining Sporadic Cancer Mutations Uncovers a Set of Genes Involved in Mitochondrial Maintenance
    (MDPI, 2023) Moreno, Armando; Taffet, Allison; Tjahjono, Elissa; Anderson, Quinton L.; Kirienko, Natalia V.
    Mitochondria are key organelles for cellular health and metabolism and the activation of programmed cell death processes. Although pathways for regulating and re-establishing mitochondrial homeostasis have been identified over the past twenty years, the consequences of disrupting genes that regulate other cellular processes, such as division and proliferation, on affecting mitochondrial function remain unclear. In this study, we leveraged insights about increased sensitivity to mitochondrial damage in certain cancers, or genes that are frequently mutated in multiple cancer types, to compile a list of candidates for study. RNAi was used to disrupt orthologous genes in the model organism Caenorhabditis elegans, and a series of assays were used to evaluate these genes’ importance for mitochondrial health. Iterative screening of ~1000 genes yielded a set of 139 genes predicted to play roles in mitochondrial maintenance or function. Bioinformatic analyses indicated that these genes are statistically interrelated. Functional validation of a sample of genes from this set indicated that disruption of each gene caused at least one phenotype consistent with mitochondrial dysfunction, including increased fragmentation of the mitochondrial network, abnormal steady-state levels of NADH or ROS, or altered oxygen consumption. Interestingly, RNAi-mediated knockdown of these genes often also exacerbated α-synuclein aggregation in a C. elegans model of Parkinson’s disease. Additionally, human orthologs of the gene set showed enrichment for roles in human disorders. This gene set provides a foundation for identifying new mechanisms that support mitochondrial and cellular homeostasis.
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    DNA supercoiling-induced shapes alter minicircle hydrodynamic properties
    (Oxford University Press, 2023) Waszkiewicz, Radost; Ranasinghe, Maduni; Fogg, Jonathan M; Catanese, Daniel J, Jr.; Ekiel-Jeżewska, Maria L; Lisicki, Maciej; Demeler, Borries; Zechiedrich, Lynn; Szymczak, Piotr
    DNA in cells is organized in negatively supercoiled loops. The resulting torsional and bending strain allows DNA to adopt a surprisingly wide variety of 3-D shapes. This interplay between negative supercoiling, looping, and shape influences how DNA is stored, replicated, transcribed, repaired, and likely every other aspect of DNA activity. To understand the consequences of negative supercoiling and curvature on the hydrodynamic properties of DNA, we submitted 336 bp and 672 bp DNA minicircles to analytical ultracentrifugation (AUC). We found that the diffusion coefficient, sedimentation coefficient, and the DNA hydrodynamic radius strongly depended on circularity, loop length, and degree of negative supercoiling. Because AUC cannot ascertain shape beyond degree of non-globularity, we applied linear elasticity theory to predict DNA shapes, and combined these with hydrodynamic calculations to interpret the AUC data, with reasonable agreement between theory and experiment. These complementary approaches, together with earlier electron cryotomography data, provide a framework for understanding and predicting the effects of supercoiling on the shape and hydrodynamic properties of DNA.
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    Consumer feces impact coral health in guild-specific ways
    (Frontiers Media S.A., 2023) Grupstra, Carsten G. B.; Howe-Kerr, Lauren I.; van der Meulen, Jesse A.; Veglia, Alex J.; Coy, Samantha R.; Correa, Adrienne M. S.
    Animal waste products are an important component of nutrient cycles and result in the trophic transmission of diverse microorganisms. There is growing recognition that the feces of consumers, such as predators, may impact resource species, their prey, via physical effects and/or microbial activity. We tested the effect of feces from distinct fish trophic groups on coral health and used heat-killed fecal controls to tease apart physical versus microbial effects of contact with fecal material. Fresh grazer/detritivore fish feces caused lesions more frequently on corals, and lesions were 4.2-fold larger than those from sterilized grazer/detritivore feces; in contrast, fresh corallivore feces did not cause more frequent or larger lesions than sterilized corallivore feces. Thus, microbial activity in grazer/detritivore feces, but not corallivore feces, was harmful to corals. Characterization of bacterial diversity in feces of 10 reef fish species, ranging from obligate corallivores to grazer/detritivores, indicated that our experimental findings may be broadly generalizable to consumer guild, since feces of some obligate corallivores contained ~2-fold higher relative abundances of coral mutualist bacteria (e.g., Endozoicomonadaceae), and lower abundances of the coral pathogen, Vibrio coralliilyticus, than feces of some grazer/detritivores. These findings recontextualize the ecological roles of consumers on coral reefs: although grazer/detritivores support coral reef health in various ways (e.g., promoting coral settlement and herbivory through the removal of detritus and sediments from the algal matrix), they also disperse coral pathogens. Corallivore predation can wound corals, yet their feces contain potentially beneficial coral-associated bacteria, supporting the hypothesized role of consumers, and corallivores in particular, in coral symbiont dispersal. Such consumer-mediated microbial dispersal as demonstrated here has broad implications for environmental management.
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    Bacterial and fungal inhibitor interacted impacting growth of invasive Triadica sebifera and soil N2O emissions
    (Frontiers Media S.A., 2023) Lai, Xiaoqin; Luo, Laicong; Fang, Haifu; Zhang, Ling; Shad, Nasir; Bai, Jian; Li, Aixin; Zhang, Xi; Yu, Yadi; Wang, Hao; Siemann, Evan
    Plant invasions affect biodiversity and seriously endanger the stability of ecosystems. Invasive plants show strong adaptability and growth advantages but are influenced by various factors. Soil bacteria and fungi are critical to plant growth and are important factors affecting plant invasions. Plant invasions also affect soil N2O emissions, but the effects of invasive plants from different population origins on N2O emissions and their microbial mechanisms are not clear. In this experiment, we grew Triadica sebifera from native (China) and invasive (USA) populations with or without bacterial (streptomycin) and/or fungal (iprodione) inhibitors in a factorial experiment in which we measured plant growth and soil N2O emissions of T. sebifera. Plants from invasive populations had higher leaf masses than those from native populations when soil bacteria were not inhibited (with or without fungal inhibition) which might reflect that they are more dependent on soil bacteria. Cumulative N2O emissions were higher for soils with invasive T. sebifera than those with a plant from a native population. Bacterial inhibitor application reduced cumulative N2O emissions but reductions were larger with application of the fungal inhibitor either alone or in combination with the bacterial inhibitor. This suggests that fungi play a strong role in plant performance and soil N2O emissions. Therefore, it is important to further understand the effects of soil microorganisms on the growth of T. sebifera and soil N2O emissions to provide a more comprehensive scientific basis for understanding the causes and consequences of plant invasions.
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    Impact of a natural disturbance on the performance and microbial communities in a full-scale constructed wetland for industrial wastewater treatment
    (Frontiers Media S.A., 2023) Hollstein, Marielle; Comerford, Mattheau; Uhl, Michael; Abel, Michael; Egan, Scott P.; Stadler, Lauren B.
    Constructed Wetlands (CWs) are a cost-effective, versatile and sustainable choice for wastewater treatment. In these environments, microbial communities play a significant role in pollutant removal. However, little is known about how microbial communities in full-scale CWs contribute to maintaining water quality or how their dynamics change in response to pulse disturbances such as fire or freezes. Furthermore, few studies have examined the relationship between CW microbial community structure and performance in full-scale industrial operations. We characterized the water-column and leaf-litter layer microbial communities in a 110-acre free water surface CW that provides tertiary wastewater treatment to a plastics manufacturing plant. The CW’s sampling campaign was conducted over a 12-month period that included Winter Storm Uri, a 100-year freeze event. Analysis of 16S rRNA gene amplicon sequences revealed that the bacterial communities experienced a temporal shift. There was also a shift in microbial community structure between the influent and the first segment of the CW. However, no differences in microbial community structure were observed in the second segment of the CW. There was a negative association between microbial community diversity and chlorophyll a, as well as microbial community diversity and total suspended solids (TSS); demonstrating an increase in microbial biodiversity as water quality improved throughout the CW. Six months after the freeze, CW performance in terms of removal of water quality constituents began to return to former removal trends. Yet, there was still a significant difference in microbial community structure within the CW relative to the previous year. This suggests CW functional resilience despite a shift in microbial community structure in the wetland.
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    pYtags enable spatiotemporal measurements of receptor tyrosine kinase signaling in living cells
    (eLife Sciences Publications Ltd., 2023) Farahani, Payam E; Yang, Xiaoyu; Mesev, Emily V; Fomby, Kaylan A; Brumbaugh-Reed, Ellen H; Bashor, Caleb J; Nelson, Celeste M; Toettcher, Jared E
    Receptor tyrosine kinases (RTKs) are major signaling hubs in metazoans, playing crucial roles in cell proliferation, migration, and differentiation. However, few tools are available to measure the activity of a specific RTK in individual living cells. Here, we present pYtags, a modular approach for monitoring the activity of a user-defined RTK by live-cell microscopy. pYtags consist of an RTK modified with a tyrosine activation motif that, when phosphorylated, recruits a fluorescently labeled tandem SH2 domain with high specificity. We show that pYtags enable the monitoring of a specific RTK on seconds-to-minutes time scales and across subcellular and multicellular length scales. Using a pYtag biosensor for epidermal growth factor receptor (EGFR), we quantitatively characterize how signaling dynamics vary with the identity and dose of activating ligand. We show that orthogonal pYtags can be used to monitor the dynamics of EGFR and ErbB2 activity in the same cell, revealing distinct phases of activation for each RTK. The specificity and modularity of pYtags open the door to robust biosensors of multiple tyrosine kinases and may enable engineering of synthetic receptors with orthogonal response programs.
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    Pathogen-induced dormancy in liquid limits gastrointestinal colonization of Caenorhabditis elegans
    (Taylor & Francis, 2023) Zhang, Liyang; Gade, Vyshnavi; Kirienko, Natalia V.
    Colonization is generally considered a prerequisite for infection, but this event is context-dependent, as evidenced by the differing ability of the human pathogen Pseudomonas aeruginosa to efficiently colonize Caenorhabditis elegans on agar but not in liquid . In this study, we examined the impact of the environment, pathogen, host, and their interactions on host colonization. We found that the transition to a liquid environment reduces food uptake by about two-fold. Also expression of specific adhesins was significantly altered in liquid-based assays for P. aeruginosa, suggesting that it may be one factor driving diminished colonization. Unexpectedly, host immune pathways did not appear to play a significant role in decreased colonization in liquid. Although knocking down key immune pathways (e.g. daf-16 or zip-2), either alone or in combination, significantly reduced survival, the changes in colonization were very small. In spite of the limited bacterial accumulation in the liquid setting, pathogenic colonization was still required for the virulence of Enterococcus faecalis. In addition, we found that a pathogen-induced dormancy was displayed by C. elegans in liquid medium after pathogen exposure, resulting in cessation of pharyngeal pumping and a decrease in bacterial intake. We conclude that poor colonization in liquid is likely due to a combination of environmental factors and host-pathogen interactions. These results provide new insights into mechanisms for colonization in different models, enabling pathogenesis models to be fine-tuned to more accurately represent the conditions seen in human infections so that new tools for curbing bacterial and fungal infections can be developed.
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    Indirect Enrichment of Desirable, but Less Fit Phenotypes, from a Synthetic Microbial Community Using Microdroplet Confinement
    (American Chemical Society, 2023) Prabhakar, Ramya Ganiga; Fan, Gaoyang; Alnahhas, Razan N.; Hirning, Andrew J.; Bennett, Matthew R.; Shamoo, Yousif
    Spatial structure within microbial communities can provide nearly limitless opportunities for social interactions and are an important driver for evolution. As metabolites are often molecular signals, metabolite diffusion within microbial communities can affect the composition and dynamics of the community in a manner that can be challenging to deconstruct. We used encapsulation of a synthetic microbial community within microdroplets to investigate the effects of spatial structure and metabolite diffusion on population dynamics and to examine the effects of cheating by one member of the community. The synthetic community was composed of three strains: a “Producer” that makes the diffusible quorum sensing molecule (N-(3-oxododecanoyl)-l-homoserine lactone, C12-oxo-HSL) or AHL; a “Receiver” that is killed by AHL; and a Non-Producer or “cheater” that benefits from the extinction of the Receivers, but without the costs associated with the AHL synthesis. We demonstrate that despite rapid diffusion of AHL between microdroplets, the spatial structure imposed by the microdroplets allows a more efficient but transient enrichment of more rare and slower-growing Producer subpopulations. Eventually, the Non-Producer population drove the Producers to extinction. By including fluorescence-activated microdroplet sorting and providing sustained competition by the Receiver strain, we demonstrate a strategy for indirect enrichment of a rare and unlabeled Producer. The ability to screen and enrich metabolite Producers from a much larger population under conditions of rapid diffusion provides an important framework for the development of applications in synthetic ecology and biotechnology.
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    Native diversity contributes to composition heterogeneity of exotic floras
    (Wiley, 2023) Chen, Pengdong; Shen, Changchao; Ren, Jie; Qin, Wenchao; Yi, Jiahui; Guan, Shuping; Tao, Zhibin; Huang, Wei; Siemann, Evan
    Variation in species composition among sites (beta diversity) is generally thought to be driven by environmental filtering and dispersal limitation, but the role of biotic interactions has not been sufficiently addressed. Specifically, the early species in a local community may contribute to subsequent beta diversity patterns. Exotic assemblages within native communities provide a unique opportunity to study biotic interaction mechanisms. In this study, we conducted a field survey of plants over an approximately 1800-km transect in the middle and lower Yangtze River valley in China to study how native communities influence exotic beta diversity. The survey included 459 plots in 51 local plant communities with 40 exotic species and 103 co-occurring native species. We also investigated how 11 environmental factors involving climate conditions, soil properties, and human activity regulate the interaction between native and exotic plants. The results showed that native diversity (Shannon–Wiener index) increased exotic beta diversity. Environmental conditions, especially monthly minimum temperature, influenced exotic beta diversity indirectly through native diversity rather than directly. Our results suggest that lower native diversity driven by environmental conditions, especially warmer temperatures, led to a decrease in composition heterogeneity of the exotic flora. Our results will help to incorporate biotic interactions into the framework of beta diversity mechanisms for local community assembly.
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    Visualization of RNA virus infection in a marine protist with a universal biomarker
    (Springer Nature, 2023) Coy, Samantha R.; Utama, Budi; Spurlin, James W.; Kim, Julia G.; Deshmukh, Harshavardhan; Lwigale, Peter; Nagasaki, Keizo; Correa, Adrienne M. S.
    Half of the marine virosphere is hypothesized to be RNA viruses (kingdom Orthornavirae) that infect abundant micro-eukaryotic hosts (e.g. protists). To test this, quantitative approaches that broadly track infections in situ are needed. Here, we describe a technique—dsRNA-Immunofluorescence (dsRIF)—that uses a double-stranded RNA (dsRNA) targeting monoclonal antibody to assess host infection status based on the presence of dsRNA, a replicative intermediate of all Orthornavirae infections. We show that the dinoflagellate Heterocapsa circularisquama produces dsRIF signal ~ 1000 times above background autofluorescence when infected by the + ssRNA virus HcRNAV. dsRNA-positive virocells were detected across > 50% of the 48-h infection cycle and accumulated to represent at least 63% of the population. Photosynthetic and chromosomal integrity remained intact during peak replication, indicating HcRNAV infection does not interrupt these processes. This work validates the use of dsRIF on marine RNA viruses and their hosts, setting the stage for quantitative environmental applications that will accelerate understanding of virus-driven ecosystem impacts.
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    Molecular Basis of KAT2A Selecting Acyl-CoA Cofactors for Histone Modifications
    (AAAS, 2023) Li, Sha; Li, Nan; He, Jie; Zhou, Runxin; Lu, Zhimin; Tao, Yizhi Jane; Guo, Yusong R.; Wang, Yugang
    Emerging discoveries about undocumented acyltransferase activities of known histone acetyltransferases (HATs) advance our understandings in the regulation of histone modifications. However, the molecular basis of HATs selecting acyl coenzyme A (acyl-CoA) substrates for histone modification is less known. We here report that lysine acetyltransferase 2A (KAT2A) as an illustrative instance of HATs can selectively utilize acetyl-CoA, propionyl-CoA, butyryl-CoA, and succinyl-CoA to directly deposit 18 histone acylation hallmarks in nucleosome. By analyzing the co-crystal structures of the catalytic domain of KAT2A in complex with acetyl-CoA, propionyl-CoA, butyryl-CoA, malonyl-CoA, succinyl-CoA, and glutaryl-CoA, we conclude that the alternative substrate-binding pocket of KAT2A and the length and electrostatic features of the acyl chain cooperatively determine the selection of the acyl-CoA substrates by KAT2A. This study reveals the molecular basis underlying the pluripotency of HATs that selectively install acylation hallmarks in nucleosomes, which might serve as instrumental mechanism to precisely regulate histone acylation profiles in cells.
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    An associative memory Hamiltonian model for DNA and nucleosomes
    (PLOS, 2023) Lu, Weiqi; Onuchic, José N.; Pierro, Michele Di; Center for Theoretical Biological Physics
    A model for DNA and nucleosomes is introduced with the goal of studying chromosomes from a single base level all the way to higher-order chromatin structures. This model, dubbed the Widely Editable Chromatin Model (WEChroM), reproduces the complex mechanics of the double helix including its bending persistence length and twisting persistence length, and the temperature dependence of the former. The WEChroM Hamiltonian is composed of chain connectivity, steric interactions, and associative memory terms representing all remaining interactions leading to the structure, dynamics, and mechanical characteristics of the B-DNA. Several applications of this model are discussed to demonstrate its applicability. WEChroM is used to investigate the behavior of circular DNA in the presence of positive and negative supercoiling. We show that it recapitulates the formation of plectonemes and of structural defects that relax mechanical stress. The model spontaneously manifests an asymmetric behavior with respect to positive or negative supercoiling, similar to what was previously observed in experiments. Additionally, we show that the associative memory Hamiltonian is also capable of reproducing the free energy of partial DNA unwrapping from nucleosomes. WEChroM is designed to emulate the continuously variable mechanical properties of the 10nm fiber and, by virtue of its simplicity, is ready to be scaled up to molecular systems large enough to investigate the structural ensembles of genes. WEChroM is implemented in the OpenMM simulation toolkits and is freely available for public use.
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    A discrete intermediate for the biosynthesis of both the enediyne core and the anthraquinone moiety of enediyne natural products
    (PNAS, 2023) Bhardwaj, Minakshi; Cui, Zheng; Daniel Hankore, Erome; Moonschi, Faruk H.; Saghaeiannejad Esfahani, Hoda; Kalkreuter, Edward; Gui, Chun; Yang, Dong; Phillips, George N.; Thorson, Jon S.; Shen, Ben; Van Lanen, Steven G.
    The enediynes are structurally characterized by a 1,5-diyne-3-ene motif within a 9- or 10-membered enediyne core. The anthraquinone-fused enediynes (AFEs) are a subclass of 10-membered enediynes that contain an anthraquinone moiety fused to the enediyne core as exemplified by dynemicins and tiancimycins. A conserved iterative type I polyketide synthase (PKSE) is known to initiate the biosynthesis of all enediyne cores, and evidence has recently been reported to suggest that the anthraquinone moiety also originates from the PKSE product. However, the identity of the PKSE product that is converted to the enediyne core or anthraquinone moiety has not been established. Here, we report the utilization of recombinant E. coli coexpressing various combinations of genes that encode a PKSE and a thioesterase (TE) from either 9- or 10-membered enediyne biosynthetic gene clusters to chemically complement ΔPKSE mutant strains of the producers of dynemicins and tiancimycins. Additionally, 13C-labeling experiments were performed to track the fate of the PKSE/TE product in the ΔPKSE mutants. These studies reveal that 1,3,5,7,9,11,13-pentadecaheptaene is the nascent, discrete product of the PKSE/TE that is converted to the enediyne core. Furthermore, a second molecule of 1,3,5,7,9,11,13-pentadecaheptaene is demonstrated to serve as the precursor of the anthraquinone moiety. The results establish a unified biosynthetic paradigm for AFEs, solidify an unprecedented biosynthetic logic for aromatic polyketides, and have implications for the biosynthesis of not only AFEs but all enediynes.
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    Primer terminal ribonucleotide alters the active site dynamics of DNA polymerase η and reduces DNA synthesis fidelity
    (Elsevier, 2023) Chang, Caleb; Lee Luo, Christie; Eleraky, Sarah; Lin, Aaron; Zhou, Grace; Gao, Yang
    DNA polymerases catalyze DNA synthesis with high efficiency, which is essential for all life. Extensive kinetic and structural efforts have been executed in exploring mechanisms of DNA polymerases, surrounding their kinetic pathway, catalytic mechanisms, and factors that dictate polymerase fidelity. Recent time-resolved crystallography studies on DNA polymerase η (Pol η) and β have revealed essential transient events during the DNA synthesis reaction, such as mechanisms of primer deprotonation, separated roles of the three metal ions, and conformational changes that disfavor incorporation of the incorrect substrate. DNA-embedded ribonucleotides (rNs) are the most common lesion on DNA and a major threat to genome integrity. While kinetics of rN incorporation has been explored and structural studies have revealed that DNA polymerases have a steric gate that destabilizes ribonucleotide triphosphate binding, the mechanism of extension upon rN addition remains poorly characterized. Using steady-state kinetics, static and time-resolved X-ray crystallography with Pol η as a model system, we showed that the extra hydroxyl group on the primer terminus does alter the dynamics of the polymerase active site as well as the catalysis and fidelity of DNA synthesis. During rN extension, Pol η error incorporation efficiency increases significantly across different sequence contexts. Finally, our systematic structural studies suggest that the rN at the primer end improves primer alignment and reduces barriers in C2′-endo to C3′-endo sugar conformational change. Overall, our work provides further mechanistic insights into the effects of rN incorporation on DNA synthesis.