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At least 91 records · Page 5

The cis-regulatory codes of response to combined heat and drought stress in Arabidopsis thaliana

Abstract Plants respond to their environment by dynamically modulating gene expression. A powerful approach for understanding how these responses are regulated is to integrate information about cis-regulatory elements (CREs) into models called cis-regulatory codes. Transcriptional response to combined stress is typically not the sum of the responses to the individual stresses. However, cis-regulatory codes underlying combined stress response have not been established. Here we modeled transcriptional response to single and combined heat and drought stress in Arabidopsis thaliana. We grouped genes by their pattern of response (independent, antagonistic and synergistic) and trained machine learning models to predict their response using putative CREs (pCREs) as features (median F-measure = 0.64). We then developed a deep learning approach to integrate additional omics information (sequence conservation, chromatin accessibility and histone modification) into our models, improving performance by 6.2%. While pCREs important for predicting independent and antagonistic responses tended to resemble binding motifs of transcription factors associated with heat and/or drought stress, important synergistic pCREs resembled binding motifs of transcription factors not known to be associated with stress. These findings demonstrate how in silico approaches can improve our understanding of the complex codes regulating response to combined stress and help us identify prime targets for future characterization.

Azodi, Christina B.↗

Detection of trans–cis flips and peptide-plane flips in protein structures

A method is presented to detect peptide bonds that need either a trans–cis flip or a peptide-plane flip. A coordinate-based method is presented to detect peptide bonds that need correction either by a peptide-plane flip or by a trans–cis inversion of the peptide bond. When applied to the whole Protein Data Bank, the method predicts 4617 trans–cis flips and many thousands of hitherto unknown peptide-plane flips. A few examples are highlighted for which a correction of the peptide-plane geometry leads to a correction of the understanding of the structure–function relation. All data, including 1088 manually validated cases, are freely available and the method is available from a web server, a web-service interface and through WHAT-CHECK.

71 CLASSICAL AND QUANTUM MECHANICS, GENERAL PHYSIC↗

Engineering quantitative stomatal trait variation and local adaptation potential by cis‐regulatory editing

Summary Cis‐regulatory element editing can generate quantitative trait variation that mitigates extreme phenotypes and harmful pleiotropy associated with coding sequence mutations. Here, we applied a multiplexed CRISPR/Cas9 approach, informed by bioinformatic datasets, to generate genotypic variation in the promoter ofOsSTOMAGEN, a positive regulator of rice stomatal density. Engineered genotypic variation corresponded to broad and continuous variation in stomatal density, ranging from 70% to 120% of wild‐type stomatal density. This panel of stomatal variants was leveraged in physiological assays to establish discrete relationships between stomatal morphological variation and stomatal conductance, carbon assimilation and intrinsic water use efficiency in steady‐state and fluctuating light conditions. Additionally, promoter alleles were subjected to vegetative drought regimes to assay the effects of the edited alleles on developmental response to drought. Notably, the capacity for drought‐responsive stomatal density reprogramming instomagenand two cis‐regulatory edited alleles was reduced. Collectively our data demonstrate that cis‐regulatory element editing can generate near‐isogenic trait variation that can be leveraged for establishing relationships between anatomy and physiology, providing a basis for optimizing traits across diverse environments.

Biotechnology & Applied Microbiology↗

Comparative genomics of metabolic capacities of regulons controlled by cis-regulatory RNA motifs in bacteria

Background: In silico comparative genomics approaches have been efficiently used for functional prediction and reconstruction of metabolic and regulatory networks. Riboswitches are metabolite-sensing structures often found in bacterial mRNA leaders controlling gene expression on transcriptional or translational levels. An increasing number of riboswitches and other cis-regulatory RNAs have been recently classified into numerous RNA families in the Rfam database. High conservation of these RNA motifs provides a unique advantage for their genomic identification and comparative analysis. Results: A comparative genomics approach implemented in the RegPredict tool was used for reconstruction and functional annotation of regulons controlled by RNAs from 43 Rfam families in diverse taxonomic groups of Bacteria. The inferred regulons include ~5200 cis-regulatory RNAs and more than 12000 target genes in 255 microbial genomes. All predicted RNA-regulated genes were classified into specific and overall functional categories. Analysis of taxonomic distribution of these categories allowed us to establish major functional preferences for each analyzed cis-regulatory RNA motif family. Overall, most RNA motif regulons showed predictable functional content in accordance with their experimentally established effector ligands. Our results suggest that some RNA motifs (including thiamin pyrophosphate and cobalamin riboswitches that control the cofactor metabolism) are widespread and likely originated from the last common ancestor of all bacteria. However, many more analyzed RNA motifs are restricted to a narrow taxonomic group of bacteria and likely represent more recent evolutionary innovations. Conclusions: The reconstructed regulatory networks for major known RNA motifs substantially expand the existing knowledge of transcriptional regulation in bacteria. The inferred regulons can be used for genetic experiments, functional annotations of genes, metabolic reconstruction and evolutionary analysis. The obtained genome-wide collection of reference RNA motif regulons is available in the RegPrecise database (http://regprecise.lbl.gov/).

59 BASIC BIOLOGICAL SCIENCES↗

Chemiluminescence Study of the Autoxidation of cis-1,4-Polyisoprene

The free-radical mechanism for the autoxidation of cis-1,4-polyisoprene (natural rubber or its synthetic counterpart) has been investigated extensively. An important feature of this mechanism, and indeed also of the autoxidation of hydrocarbons generally, is that it is a chain process propagated by alkyl and peroxy radicals and terminated through bimolecular reactions involving these same radicals. In the usual oxidation situation, that is, at all oxygen pressures greater than a few torr, the alkyl radicals are rapidly converted to peroxy radicals, and the termination step proceeds almost exclusively through the latter radicals. The bimolecular decay of the peroxy radicals is accompanied by a weak emission of light or chemiluminescence. Kinetic evidence is consistent with an electronically excited ketone produced in the termination reaction as the source of the emission. The first observation of chemiluminescence from the oxidative degradation of polymers was reported by Ashby, who dealt mainly with polypropylene but made passing mention of several other polymers. Subsequently, a number of papers have appeared dealing with oxidative chemiluminescence from a variety of polymers. In this paper we report the first detailed study of the chemiluminescence emitted in the autoxidation of cis-1,4-polyisoprene. The chemiluminescence technique is extremely sensitive and can follow rates of oxidation that are too slow to be measured conveniently by other means. This work thus offered the potential of throwing new light on the autoxidation of cis-1,4-polyisoprene, especially in the very early stages or under ambient conditions where conventional spectroscopic procedures are rather insensitive.

Mendenhall, G. David↗

A Cis-Lunar Propellant Infrastructure for Flexible Path Exploration and Space Commerce

This paper describes a space infrastructure concept that exploits lunar water for propellant production and delivers it to users in cis-lunar space. The goal is to provide responsive economical space transportation to destinations beyond low Earth orbit (LEO) and enable in-space commerce. This is a game changing concept that could fundamentally affect future space operations, provide greater access to space beyond LEO, and broaden participation in space exploration. The challenge is to minimize infrastructure development cost while achieving a low operational cost. This study discusses the evolutionary development of the infrastructure from a very modest robotic operation to one that is capable of supporting human operations. The cis-lunar infrastructure involves a mix of technologies including cryogenic propellant production, reusable lunar landers, propellant tankers, orbital transfer vehicles, aerobraking technologies, and electric propulsion. This cislunar propellant infrastructure replaces Earth-launched propellants for missions beyond LEO. It enables users to reach destinations with smaller launchers or effectively multiplies the user s existing payload capacity. Users can exploit the expanded capacity to launch logistics material that can then be traded with the infrastructure for propellants. This mutually beneficial trade between the cis-lunar infrastructure and propellant users forms the basis of in-space commerce.

Oeftering, Richard C.↗

Low Thrust Cis-Lunar Transfers Using a 40 kW-Class Solar Electric Propulsion Spacecraft

This paper captures trajectory analysis of a representative low thrust, high power Solar Electric Propulsion (SEP) vehicle to move a mass around cis-lunar space in the range of 20 to 40 kW power to the Electric Propulsion (EP) system. These cis-lunar transfers depart from a selected Near Rectilinear Halo Orbit (NRHO) and target other cis-lunar orbits. The NRHO cannot be characterized in the classical two-body dynamics more familiar in the human spaceflight community, and the use of low thrust orbit transfers provides unique analysis challenges. Among the target orbit destinations documented in this paper are transfers between a Southern and Northern NRHO, transfers between the NRHO and a Distant Retrograde Orbit (DRO) and a transfer between the NRHO and two different Earth Moon Lagrange Point 2 (EML2) Halo orbits. Because many different NRHOs and EML2 halo orbits exist, simplifying assumptions rely on previous analysis of orbits that meet current abort and communication requirements for human mission planning. Investigation is done into the sensitivities of these low thrust transfers to EP system power. Additionally, the impact of the Thrust to Weight ratio of these low thrust SEP systems and the ability to transit between these unique orbits are investigated.

cislunar↗

The biosynthesis of the anti–microbial diterpenoid leubethanol in Leucophyllum frutescens proceeds via an all– cis prenyl intermediate

Serrulatane diterpenoids are natural products found in plants from a subset of genera within the figwort family (Scrophulariaceae). Many of these compounds have been characterized as having anti–microbial properties and share a common diterpene backbone. One example, leubethanol from Texas sage (Leucophyllum frutescens) has demonstrated activity against multi–drug–resistant tuberculosis. Leubethanol is the only serrulatane diterpenoid identified from this genus; however, a range of such compounds have been found throughout the closely related Eremophila genus. Despite their potential therapeutic relevance, the biosynthesis of serrulatane diterpenoids has not been previously reported. Here we leverage the simple product profile and high accumulation of leubethanol in the roots of L. frutescens and compare tissue–specific transcriptomes with existing data from Eremophila serrulata to decipher the biosynthesis of leubethanol. A short–chain cis–prenyl transferase (LfCPT1) first produces the rare diterpene precursor nerylneryl diphosphate, which is cyclized by an unusual plastidial terpene synthase (LfTPS1) into the characteristic serrulatane diterpene backbone. Final conversion to leubethanol is catalyzed by a cytochrome P450 (CYP71D616) of the CYP71 clan. This pathway documents the presence of a short–chain cis–prenyl diphosphate synthase, previously only found in Solanaceae, which is likely involved in the biosynthesis of other known diterpene backbones in Eremophila. LfTPS1 represents neofunctionalization of a compartment–switching terpene synthase accepting a novel substrate in the plastid. Biosynthetic access to leubethanol will enable pathway discovery to more complex serrulatane diterpenoids which share this common starting structure and provide a platform for the production and diversification of this class of promising anti–microbial therapeutics in heterologous systems.

59 BASIC BIOLOGICAL SCIENCES↗

On the morphological behavior of ABC miktoarm stars containing poly(cis 1,4-isoprene), poly(styrene), and poly(2-vinylpyridine)

Fundamental understanding of microphase separation in ABC miktoarm copolymers is vital to access a plethora of nonconventional morphologies. Miktoarm stars based on poly(cis 1,4–isoprene) (I), poly(styrene) (S), and poly(2–vinylpyridine) (V) are model systems, which allow systematic studies of the effects of composition, chemical microstructure, and temperature on the thermodynamics of microphase separation. Eleven ISV–x (I:S:V = 1:1:x, v:v:v) miktoarm copolymers were synthesized by anionic polymerization affording well-defined copolymers with a variable V arm. Equilibrium bulk morphologies of all samples, as evidenced by small–angle X–ray scattering, transmission electron microscopy (TEM), and self–consistent field theory, showed a systematic transition from lamellae (x ≈ 0–0.2) to [8.8.4] tiling (x ≈ 0.6–0.9) to cylinders in undulating lamellae (x ≈ 2–4) and, finally, to hexagonally packed core–shell cylinders (x ≈ 5–8). Chemical microstructure of the I arm [poly(cis 1,4–isoprene)] versus poly(3,4–isoprene) is shown to play important role in affecting morphological behavior. Furthermore, to reconcile differences between ISV–x star morphologies reported in the literature and those reported herein, even for the same composition, effects of the microstructure of I arm on the Flory–Huggins parameter between I and V arms were taken into account in a qualitative manner.

36 MATERIALS SCIENCE↗

Photochemistry of cis,trans-[Pt{sup IV}(en)(I){sub 2}(CH{sub 3}COO){sub 2}] complex in aqueous solutions

The photochemistry of aqueous solutions of the cis,trans-[Pt{sup IV}(en)(I){sub 2}(CH{sub 3}COO){sub 2}] complex (1) was studied by stationary photolysis, nanosecond laser flash photolysis and ultrafast kinetic spectroscopy in the time interval from hundreds of femtoseconds to minutes. It is shown that the multistage photolysis, including photochemical reactions of ligand exchange and photoreduction, involves in the fi rst step the formation of the cis-[{sup IV}(en)(I){sub 2}(CH{sub 3}COO){sub 2}] complex with a quantum yield of 0.3 (irradiation at 282 nm). The further ligand exchange and photoreduction reactions occur with much lower quantum yields. The photochemical exchange of the acetate ion occurs in a time interval of ~100 ps with the much slower final step, being probably the acidic dissociation of the product. The key intermediate of the first step of the process was detected in the picosecond time range. Depending on the interpretation of the key intermediate nature, two possible mechanisms of the primary photochemical process are proposed.

37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CH↗

Study of the Rotational Structure of the Forbidden ν{sub 8} Band of the C{sub 2}H{sub 2}D{sub 2}-CIS Molecule

The spectrum of the cis-ethylene-d2 (C{sub 2}H{sub 2}D{sub 2}-cis) molecule is recorded with a Bruker IFS 120 HR Fourier spectrometer in the region of 600–1200 cm{sup –1} with spectral resolution of 0.0021 cm{sup –1}. As a result of analysis of the experimental spectrum, 36 transitions are first assigned, based on which seven upper energy levels of the state (ν{sub 8} = 1) are determined. The ν{sub 8} band is symmetry-forbidden, and its transitions appear in the IR spectrum only due to the strong c-type Coriolis interaction between the states (ν{sub 8} = 1) and (ν{sub 7} = 1). The parameters describing the rotational structure of the ν{sub 8} band are obtained.

74 ATOMIC AND MOLECULAR PHYSICS↗

Substance P in Solution: Trans-to-Cis Configurational Changes of Penultimate Prolines Initiate Non-enzymatic Peptide Bond Cleavages

We report ion mobility spectrometry and mass spectrometry studies of the non-enzymatic step-by-step degradation of substance P (subP), an 11-residue neuropeptide, with the sequence Arg{sup 1}-Pro{sup 2}-Lys{sup 3}-Pro{sup 4}-Gln{sup 5}-Gln{sup 6}-Phe{sup 7}-Phe{sup 8}-Gly{sup 9}-Leu{sup 10}-Met{sup 11}-NH{sub 2}, in ethanol. At elevated solution temperatures (55 to 75 °C), several reactions are observed, including a protonation event, i.e., [subP+2H]{sup 2+} + H{sup +} → [subP+3H]{sup 3+}, that appears to be regulated by a configurational change and two sequential bond cleavages (the Pro{sup 2}-Lys{sup 3} peptide bond is cleaved to form the smaller nonapeptide Lys{sup 3}-Met{sup 11}-NH{sub 2} [subP{sub (3–11)}], and subsequently, subP{sub (3–11)} is cleaved at the Pro{sup 4}-Gln{sup 5} peptide bond to yield the heptapeptide Gln{sup 5}-Met{sup 11}-NH{sub 2} [subP{sub (5–11)}]). Each of the product peptides [subP{sub (3–11)} and subP{sub (5–11)}] is accompanied by a complementary diketopiperazine (DKP): cyclo-Arg{sup 1}-Pro{sup 2} (cRP) for the first cleavage, and cyclo-Lys{sup 3}-Pro{sup 4} (cKP) for the second. Insight about the mechanism of degradation is obtained by comparing kinetics calculations of trial model mechanisms with experimental data. The best model of our experimental data indicates that the initial cleavage of subP is regulated by a conformational change, likely a trans→cis isomerization of the Arg{sup 1}-Pro{sup 2} peptide bond. The subP{sub (3–11)} product has a long lifetime (t{sub 1/2} ~ 30 h at 55 °C) and appears to transition through several structural intermediates prior to dissociation, suggesting that subP{sub (3–11)} is initially formed with a Lys{sup 3}-trans-Pro{sup 4} peptide bond configuration and that slow trans→cis isomerization regulates the second bond cleavage event as well. From these data and our model mechanisms, we obtain transition state thermochemistry ranging from ΔH{sup ‡} = 41 to 85 kJ mol{sup −1} and ΔS{sup ‡} = − 43 to – 157 J mol{sup −1} K{sup −1} for each step in the reaction. .

37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CH↗

Combinatorial transcription factor binding encodes cis -regulatory wiring of mouse forebrain GABAergic neurogenesis

Transcription factors (TFs) bind combinatorially to cis-regulatory elements, orchestrating transcriptional programs. Although studies of chromatin state and chromosomal interactions have demonstrated dynamic neurodevelopmental cis-regulatory landscapes, parallel understanding of TF interactions lags. To elucidate combinatorial TF binding driving mouse basal ganglia development, we integrated chromatin immunoprecipitation sequencing (ChIP-seq) for twelve TFs, H3K4me3-associated enhancer-promoter interactions, chromatin and gene expression data, and functional enhancer assays. We identified sets of putative regulatory elements with shared TF binding (TF-pRE modules) that orchestrate distinct processes of GABAergic neurogenesis and suppress other cell fates. The majority of pREs were bound by one or two TFs; however, a small proportion were extensively bound. These sequences had exceptional evolutionary conservation and motif density, complex chromosomal interactions, and activity as in vivo enhancers. Our results provide insights into the combinatorial TF-pRE interactions that activate and repress expression programs during telencephalon neurogenesis and demonstrate the value of TF binding toward modeling developmental transcriptional wiring.

59 BASIC BIOLOGICAL SCIENCES↗

Evidence for cis Amide Bonds in Peptoid Nanosheets

Peptoid nanosheets are supramolecular protein-mimetic materials that form from amphiphilic polypeptoids with aromatic and ionic side chains. Nanosheets have been studied at the nanometer scale, but the molecular structure has been difficult to probe. We introduce the use of 13 C- 13 C dipolar recoupling solid-state NMR measurements to reveal the configuration of backbone amide bonds selected by 13 C isotopic labeling of adjacent α-carbons. Measurements on the same molecules in the amorphous state and in nanosheets revealed that amide bonds in the center of the amino block of peptoid (NaeNpe) 7 -(NceNpe) 7 (B28) favor the trans configuration in the amorphous state and the cis configuration in the nanosheet. This unexpected conclusion contrasts with previous NMR and theoretical studies of short solvated peptoids. Moreover, examination of the amide bond at the junction of the two charged blocks within B28 revealed a mixture of both cis and trans configurational states, consistent with the previously predicted brickwork-like intermolecular organization.

37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CH↗

Uranium-Mediated Peroxide Activation and a Precursor toward an Elusive Uranium cis -Dioxo Fleeting Intermediate

The activation of chalcogen–chalcogen bonds using organometallic uranium complexes has been well documented for S–S, Se–Se, and Te–Te bonds. In stark contrast, reports concerning the ability of a uranium complex to activate the O–O bond of an organic peroxide are exceedingly rare. Herein, we describe the peroxide O–O bond cleavage of 9,10-diphenylanthracene-9,10-endoperoxide in nonaqueous media, mediated by a uranium(III) precursor [(( Me,Ad ArO) 3 N)U III (dme)] to generate a stable uranium(V) bis-alkoxide complex, namely, [(( Me,Ad ArO) 3 N)U V (DPAP)]. This reaction proceeds via an isolable, alkoxide-bridged diuranium(IV/IV) species, implying that the oxidative addition occurs in two sequential, single-electron oxidations of the metal center, including rebound of a terminal oxygen radical. Furthermore, this uranium(V) bis-alkoxide can then be reduced with KC 8 to form a uranium(IV) complex, which upon exposure to UV light, in solution, releases 9,10-diphenylanthracene to generate a cyclic uranyl trimer through formal two-electron photooxidation. Analysis of the mechanism of this photochemical oxidation via density functional theory (DFT) calculations indicates that the formation of this uranyl trimer occurs through a fleeting uranium cis-dioxo intermediate. At room temperature, this cis-configured dioxo species rapidly isomerizes to a more stable trans configuration through the release of one of the alkoxide ligands from the complex, which then goes on to form the isolated uranyl trimer complex.

37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CH↗

Universal Relationship between Molecular Structure and Crystal Structure in Peptoid Polymers and Prevalence of the cis Backbone Conformation

Peptoid polymers are often crystalline in the solid-state as examined by X-ray scattering, but thus far, there has been no attempt to identify a common structural motif among them. In order to probe the relationship between molecular structure and crystal structure, we synthesized and analyzed a series of crystalline peptoid copolymers, systematically varying peptoid side-chain length (S) and main-chain length (N). We also examined X-ray scattering data from 18 previously reported peptoid polymers. In all peptoids, we found that the unit cell dimensions, a, b, and c, are simple functions of S and N: a (Å) = 4.55, b (Å) = [2.98]N + 0.35, and c (Å) = [1.86]S + 5.5. These relationships, which apply to both bulk crystals and self-assembled nanosheets in water, indicate that the molecules adopt extended, planar conformations. Furthermore, we performed molecular dynamics simulations (MD) of peptoid polymer lattices, which indicate that all backbone amides adopt the cis conformation. This is a surprising conclusion, because previous studies on isolated molecules indicated an energetic preference for the trans conformer. Finally, this study demonstrates that when packed into supramolecular lattices or crystals, peptoid polymers prefer to adopt a regular, extended, all-cis secondary structure.

36 MATERIALS SCIENCE↗

Stereocontrolled Syntheses of Functionalized cis - and trans -Siladecalins

Here, we study the synthesis of both diastereomers of an all-silicon analog of decalin, the ubiquitous bicyclic structural motif. The synthesis provides materials functionalized with either Si–Ph or Si–H groups, versatile entry points for further chemical diversification. The synthesis of silicon-stereogenic silanes is significantly less precedented than the synthesis of asymmetric carbon centers and strategies for control of relative stereochemistry in oligosilanes are hardly described. This report provides insights of potential generality, such as the epimerization of the cis-isomer to the thermodynamically downhill trans-isomer via a hypothesized pentavalent intermediate. Decalin is a classic example in the conformational analysis of organic ring systems and the carbocyclic diastereomers have highly divergent conformational profiles. Like the carbocycle, we observe different conformational properties in cis- and trans-siladecalins with consequences for NMR spectroscopy, optical properties, and vibrational spectroscopy. Our research showcases the utility of targeted synthesis for preparing complex and functionalized polycyclic silanes.

36 MATERIALS SCIENCE↗

Systematic mapping of occluded genes by cell fusion reveals prevalence and stability of cis-mediated silencing in somatic cells

Both diffusible factors acting in trans and chromatin components acting in cis are implicated in gene regulation, but the extent to which either process causally determines a cell’s transcriptional identity is unclear. We recently used cell fusion to define a class of silent genes termed ‘‘cis-silenced’’ (or ‘‘occluded’’) genes, which remain silent even in the presence of transacting transcriptional activators. We further showed that occlusion of lineage-inappropriate genes plays a critical role in maintaining the transcriptional identities of somatic cells. Here, we present, for the first time, a comprehensive map of occluded genes in somatic cells. Specifically, we mapped occluded genes in mouse fibroblasts via fusion to a dozen different rat cell types followed by whole-transcriptome profiling. We found that occluded genes are highly prevalent and stable in somatic cells, representing a sizeable fraction of silent genes. Occluded genes are also highly enriched for important developmental regulators of alternative lineages, consistent with the role of occlusion in safeguarding cell identities. Alongside this map, we also present whole-genome maps of DNA methylation and eight other chromatin marks. These maps uncover a complex relationship between chromatin state and occlusion. Furthermore, we found that DNA methylation functions as the memory of occlusion in a subset of occluded genes, while histone deacetylation contributes to the implementation but not memory of occlusion. Our data suggest that the identities of individual cell types are defined largely by the occlusion status of their genomes. The comprehensive reference maps reported here provide the foundation for future studies aimed at understanding the role of occlusion in development and disease.

59 BASIC BIOLOGICAL SCIENCES↗