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At least 37 records · Page 2

A hidden cysteine in Fis1 targeted to prevent excessive mitochondrial fission and dysfunction under oxidative stress

Fis1-mediated mitochondrial localization of Drp1 and excessive mitochondrial fission occur in human pathologies associated with oxidative stress. However, it is not known how Fis1 detects oxidative stress and what structural changes in Fis1 enable mitochondrial recruitment of Drp1. We find that conformational change involving α1 helix in Fis1 exposes its only cysteine, Cys41. In the presence of oxidative stress, the exposed Cys41 in activated Fis1 forms a disulfide bridge and the Fis1 covalent homodimers cause increased mitochondrial fission through increased Drp1 recruitment to mitochondria. Our discovery of a small molecule, SP11, that binds only to activated Fis1 by engaging Cys41, and data from genetically engineered cell lines lacking Cys41 strongly suggest a role of Fis1 homodimerization in Drp1 recruitment to mitochondria and excessive mitochondrial fission. The structure of activated Fis1-SP11 complex further confirms these insights related to Cys41 being the sensor for oxidative stress. Importantly, SP11 preserves mitochondrial integrity and function in cells during oxidative stress and thus may serve as a candidate molecule for the development of treatment for diseases with underlying Fis1-mediated mitochondrial fragmentation and dysfunction.

59 BASIC BIOLOGICAL SCIENCES↗

Development and Implementation of a Nuclear and Criticality Safety Engineering Pipeline Course at North Carolina State University

Savannah River Nuclear Solutions, owner of both the criticality safety program and accident analysis qualification at Savannah River Site, experienced increasing difficulty in recruiting, training, and retaining key talent areas. In an effort to curb attrition, provide a talent base to recruit from, and introduce potential new hires to niche subject areas, a pipeline college course was developed for a regional university. The course introduces a variety of topic areas particular to criticality safety and nuclear safety at Department of Energy nonreactor nuclear facilities. Several administrative and developmental hurdles were encountered before the course was successfully initiated at North Carolina State University in fall 2024.

criticality↗

Assessment of ROI for Workforce Development Efforts National & Homeland Security

The IDEAL Professional Engagement project at Idaho National Laboratory (INL) aims to enhance the laboratory's workforce diversity and inclusivity efforts, focusing on the U.S. National and Homeland Security mission areas. This project involves researching and evaluating opportunities for INL to engage in various professional events, particularly cyber conferences, to support recruitment and professional development. The project involved several key tasks: compiling comprehensive information on national laboratories and their mission statements, developing a deep understanding of INL’s role and efforts in national and homeland security, and identifying and engaging key stakeholders. Interviews were conducted with a set of targeted questions, and the findings were analyzed to identify common themes, insights, and actionable recommendations. Additionally, relevant upcoming cyber conferences were identified, various sponsorship levels and their associated benefits were evaluated, and the recruitment potential of these conferences was assessed. A detailed cost analysis was performed, including registration fees and travel expenses, and a cost-benefit analysis was conducted to evaluate the financial viability and potential return on investment (ROI) of conference participation and sponsorship. Based on the research and analysis, actionable recommendations for conference participation and sponsorship were formulated, ensuring alignment with INL’s mission and diversity goals. Preliminary results include a comprehensive list of relevant cyber conferences, a detailed cost analysis, and a set of actionable recommendations for future conference participation and sponsorship. The analysis highlights the financial requirements and geographical distribution of these conferences, providing valuable insights for INL's engagement strategies. This project underscores the importance of strategic engagement in professional events to attract and develop a diverse and skilled workforce, ultimately supporting INL’s mission areas in U.S. National and Homeland Security.

99 GENERAL AND MISCELLANEOUS↗

Resilient information and inference networks under mixed-trust sensing

With ubiquitous digitization, sensing, and computational intelligence deployed in increasingly more and broader domains, including critical infrastructure, potentially misleading and destabilizing effects of multimodal anomalies and adversarial behavior are growing in importance. Here, we develop randomized and reinforcement learning-based strategies for strategically recruiting and utilizing deployed (and, thus, vulnerable and potentially faulty and/or compromised) nodes from information and inference networks, while defending against adversaries that attempt to misguide assessments of inferred variables. Recognizing that, besides communication and other costs, sampling from any observable node can either provide true data or dangerously expose our inference to misinformation (without being easily distinguishable what actually happens), the proposed strategies proceed by progressively recruiting nodes and cautiously scaling their information contribution based on assumed, or, in our reinforcement learning approach, intelligently weighed trustworthiness, with the learning approach also considering network-wide, threat-inclusive risk/value tradeoffs. While avoiding the hardware, communication, analytical and computational burden of explicit redundancy, the proposed defensive schemes enable on-the-fly assessments of underlying processes, and system-wide situational awareness with demonstrable resilience against adversarial activities.

97 - MATHEMATICS AND COMPUTING↗

VPS26 Moonlights as a β-Arrestin-like Adapter for a 7-Transmembrane RGS Protein in Arabidopsis thaliana

Extracellular signals perceived by 7-transmembrane (7TM)-spanning receptors initiate desensitization that involves the removal of these receptors from the plasma membrane. Agonist binding often evokes phosphorylation in the flexible C-terminal region and/or intracellular loop 3 of many 7TM G-protein-coupled receptors in animal cells, which consequently recruits a cytoplasmic intermediate adaptor, β-arrestin, resulting in clathrin-mediated endocytosis (CME) and downstream signaling such as transcriptional changes. Some 7TM receptors undergo CME without recruiting β-arrestin, but it is not clear how. Arrestins are not encoded in the Arabidopsis thaliana genome, yet Arabidopsis cells have a well-characterized signal-induced CME of a 7TM protein, designated Regulator of G Signaling 1 (AtRGS1). Here we show that a component of the retromer complex, Vacuolar Protein Sorting-Associated 26 (VPS26), binds the phosphorylated C-terminal region of AtRGS1 as a VPS26A/B heterodimer to form a complex that is required for downstream signaling. We propose that VPS26 moonlights as an arrestin-like adaptor in the CME of AtRGS1.

37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CH↗

Large Divergence of Projected High Latitude Vegetation Composition and Productivity Due To Functional Trait Uncertainty

Abstract Vegetation distribution and composition are expected to change in northern high latitudes under rapid warming, which regulates ecosystem functions but remains challenging to predict. Vegetation change arises from the interplay of chronic climate trends such as warming and transient demographic processes of recruitment, growth, competition, and mortality. Most predictive models overlooked the role of demographic dynamics controlled by plant traits. Here, we simulate vegetation dynamics at the Kougarok Hillslope site in Alaska under historical and future climates using the E3SM Land Model coupled to the Functionally Assembled Terrestrial Simulator (ELM‐FATES). To evaluate the roles of plant traits, we parameterize the model with 5,265 trait configurations representing diverse physiological and demographic strategies. Results show current modeled biomass, composition, and productivity are most sensitive to traits controlling photosynthetic capacity, carbon allocation, allometry, and phenology. Among all trait configurations, ∼5% reproduce in situ biomass and plant functional type (PFT) composition measured in 2016, that are indistinguishable from these two observed ecosystem states. Notably, these same trait configurations produce diverging biomass, composition, and productivity under future climate, where the uncertainty attributable to traits is twice the change attributable to climate change. The variation of projected productivity arises from emerging PFT composition under novel climate regimes, primarily explained by traits controlling cold‐induced mortality, recruitment, and allometry. Our findings highlight the importance and uncertainty of demographic dynamics and its interaction with climate change in shaping Arctic vegetation change. Improved model predictions will likely benefit from explicit consideration of vegetation demography and better constraints of critical traits.

54 ENVIRONMENTAL SCIENCES↗

Disentangling plant- and environment-mediated drivers of active rhizosphere bacterial community dynamics during short-term drought

Abstract Mitigating the effects of climate stress on crops is important for global food security. The microbiome associated with plant roots, the rhizobiome, can harbor beneficial microbes that alleviate stress, but the factors influencing their recruitment are unclear. We conducted a greenhouse experiment using field soil with a legacy of growing switchgrass and common bean to investigate the impact of short-term drought severity on the recruitment of active bacterial rhizobiome members. We applied 16S rRNA and 16S rRNA gene sequencing for both crops and metabolite profiling for switchgrass. We included planted and unplanted conditions to distinguish environment- versus plant-mediated rhizobiome drivers. Differences in community structure were observed between crops and between drought and watered and planted and unplanted treatments within crops. Despite crop-specific communities, drought rhizobiome dynamics were similar across the two crops. The presence of a plant more strongly explained the rhizobiome variation in bean (17%) than in switchgrass (3%), with a small effect of plant mediation during drought observed only for the bean rhizobiome. The switchgrass rhizobiome was stable despite changes in rhizosphere metabolite profiles between planted and unplanted treatments. We conclude that rhizobiome responses to short-term drought are crop-specific, with possible decoupling of plant exudation from rhizobiome responses.

59 BASIC BIOLOGICAL SCIENCES↗

A protein phosphatase 1 specific phos phatase ta rgeting p eptide (PhosTAP) to identify the PP1 phosphatome

Phosphoprotein phosphatases (PPPs) are the key serine/threonine phosphatases that regulate all essential signaling cascades. In particular, Protein Phosphatase 1 (PP1) dephosphorylates ~80% of all ser/thr phosphorylation sites. Here, we developed a phosphatase targeting peptide (PhosTAP) that binds all PP1 isoforms and does so with a stronger affinity than any other known PP1 regulator. This PhosTAP can be used as a PP1 recruitment tool for Phosphorylation Targeting Chimera (PhosTAC)-type recruitment in in vitro and cellular experiments, as well as in phosphoproteomics experiments to identify PP1-specific substrates and phosphosites. The latter is especially important to further our understanding of cellular signaling, as the identification of substrates and especially phosphosites that are targeted by specific phosphatases lags behind that of their kinase counterparts. Using PhosTAP-based proteomics, we show that, counter to our current understanding, many PP1 regulators are also substrates, that the number of residues between regulator PP1-binding and phosphosites vary significantly, and that PP1 counteracts the activities of mitotic kinases. Finally, we also found that Haspin kinase is a direct substrate of PP1 and that its PP1-dependent dephosphorylation modulates its activity during anaphase. Together, we show that PP1-specific PhosTAPs are a powerful tool for +studying PP1 activity in vitro and in cells.

Science & Technology - Other Topics↗

Structural basis for a highly conserved RNA-mediated enteroviral genome replication

Abstract Enteroviruses contain conserved RNA structures at the extreme 5′ end of their genomes that recruit essential proteins 3CD and PCBP2 to promote genome replication. However, the high-resolution structures and mechanisms of these replication-linked RNAs (REPLRs) are limited. Here, we determined the crystal structures of the coxsackievirus B3 and rhinoviruses B14 and C15 REPLRs at 1.54, 2.2 and 2.54 Å resolution, revealing a highly conserved H-type four-way junction fold with co-axially stacked sA-sD and sB-sC helices that are stabilized by a long-range A•C•U base-triple. Such conserved features observed in the crystal structures also allowed us to predict the models of several other enteroviral REPLRs using homology modeling, which generated models almost identical to the experimentally determined structures. Moreover, our structure-guided binding studies with recombinantly purified full-length human PCBP2 showed that two previously proposed binding sites, the sB-loop and 3′ spacer, reside proximally and bind a single PCBP2. Additionally, the DNA oligos complementary to the 3′ spacer, the high-affinity PCBP2 binding site, abrogated its interactions with enteroviral REPLRs, suggesting the critical roles of this single-stranded region in recruiting PCBP2 for enteroviral genome replication and illuminating the promising prospects of developing therapeutics against enteroviral infections targeting this replication platform.

Biochemistry & Molecular Biology↗

Structural and functional insights into the interaction between the bacteriophage T4 DNA processing proteins gp32 and Dda

Abstract Bacteriophage T4 is a classic model system for studying the mechanisms of DNA processing. A key protein in T4 DNA processing is the gp32 single-stranded DNA-binding protein. gp32 has two key functions: it binds cooperatively to single-stranded DNA (ssDNA) to protect it from nucleases and remove regions of secondary structure, and it recruits proteins to initiate DNA processes including replication and repair. Dda is a T4 helicase recruited by gp32, and we purified and crystallized a gp32–Dda–ssDNA complex. The low-resolution structure revealed how the C-terminus of gp32 engages Dda. Analytical ultracentrifugation analyses were consistent with the crystal structure. An optimal Dda binding peptide from the gp32 C-terminus was identified using surface plasmon resonance. The crystal structure of the Dda–peptide complex was consistent with the corresponding interaction in the gp32–Dda–ssDNA structure. A Dda-dependent DNA unwinding assay supported the structural conclusions and confirmed that the bound gp32 sequesters the ssDNA generated by Dda. The structure of the gp32–Dda–ssDNA complex, together with the known structure of the gp32 body, reveals the entire ssDNA binding surface of gp32. gp32–Dda–ssDNA complexes in the crystal are connected by the N-terminal region of one gp32 binding to an adjacent gp32, and this provides key insights into this interaction.

Biochemistry & Molecular Biology↗

Asymmetric loading of TnsE regulates Tn7 targeting of DNA replication structures

Abstract Tn7 transposable elements are known for their sophisticated target-site selection mechanisms. For the prototypical Tn7 element, dedicated transposon-encoded proteins direct insertions to either a conserved site in the chromosome or replicating DNA structures in conjugal plasmids, ensuring the vertical and horizontal spread of the element. While the pathway targeting the attTn7 site in the bacterial chromosome has been extensively studied, the pathway targeting DNA replication structures remains poorly understood. We have used an integrative structural biology approach to elucidate how the Tn7-encoded protein TnsE recognizes replication sites. Using native mass spectrometry, we found that TnsE forms 1:1 and 2:1 (TnsE:DNA) complexes on 3′-recessed DNA, with gain-of-function TnsE variants favoring the formation of 2:1 complexes. Structural characterization confirms that two TnsE molecules bind to DNA with the C-terminal domain of the protein recognizing duplex DNA, leaving the N-terminal domain to impose DNA substrate specificity and recruit the core transposition machinery. Collectively, our work is consistent with a model where TnsE-mediated target-site selection relies on the formation of an asymmetric TnsE:DNA complex to recruit the Tn7 transposase to DNA replication structures.

Biochemistry & Molecular Biology↗

CpoS-Inc interactions facilitate host cell modulation during Chlamydia trachomatis infection

ABSTRACT Chlamydia trachomatis ( C.t .), the leading bacterial cause of sexually transmitted infections, replicates within a unique intracellular compartment called the inclusion, which is modified by secreted proteins known as inclusion membrane (Inc) proteins. Here, we further characterize CpoS, an Inc protein previously shown to be critical for bacterial replication and inclusion development. We demonstrate that CpoS directly binds multiple coiled-coil region-containing Incs and engages Rab GTPases at a separate site. Notably, CpoS-InaC interactions facilitate the recruitment of select Arf GTPases to the inclusion membrane, while Rab recruitment occurs independently of these interactions. Biochemical and biophysical analyses revealed that Incs self-oligomerize to form higher-ordered structures, with CpoS adopting a tetrameric conformation resembling that of eukaryotic SNARE proteins. We propose that these assemblies serve as scaffolds to orchestrate vesicle docking, tethering, and fusion. Our findings highlight the intricate interplay between bacterial and host factors, revealing how C.t . leverages both Inc-Inc interactions and host protein engagement to manipulate vesicular trafficking and sustain infection.

Tijerina, Xavier [Department of Microbiology and I↗

2002 St. Louis Region Travel Survey

The 2002 St. Louis Household Travel Survey entailed the collection of weekday travel behavior characteristics of households residing in each of the eight counties that comprise the St. Louis region. In addition to collecting basic demographic and socioeconomic information about each household and its members, the survey documented specific characteristics of activities and trips, including the number and purpose of trips, trip duration, time of day, mode of transportation, and specifics of school- and work-related travel. The survey instruments contained three components: 1) the recruitment questionnaire, 2) the travel log, and 3) the retrieval questionnaire. In total, 7,046 households were recruited to participate in the study via telephone interview. Of these, 5,094 completed travel logs during a specific 24-hour period, and the information was retrieved from all household members, regardless of age. Demographic information for this study includes age, gender, education level, employment status, and household income.

1Hz data↗

2022 Central California Travel Study

# 2022 Central California Travel Study The 2022 Central California Travel Study collected demographic and travel pattern information to better understand household travel behavior in the state’s San Joaquin Valley region. ## Data Collection Agency The Fresno Council of Governments conducted the household travel study in collaboration with metropolitan planning organizations in Fresno, Kern, Kings, Madera, Merced, San Joaquin, Stanislaus, and Tulare counties in California. ## Survey Methodology The two-part study consisted of a survey and a travel diary. The survey gathered data on household demographic composition and typical travel behaviors. The travel diary gathered individual travel data during a specified travel period for all members of a given household. ## Travel Diary Methods Households with smartphones were encouraged to complete their travel diaries using the rMove smartphone app for up to seven consecutive days. Households without smartphones or those unwilling to participate via smartphone completed their travel diaries online (using the web-version of rMove) or by calling the survey call center. These households reported travel for one day (Tuesday, Wednesday, or Thursday). ## Recruitment Sampling Methods The majority of the recruitment was conducted via address-based sampling, a type of probability sampling, with a focus on reaching county-level targets in collaboration with metropolitan planning organizations in the region. Supplemental sampling methods, primarily non-probability, were employed during all waves of data collection to improve survey representation. The supplemental sample included targeted outreach to hard-to-survey populations via transit rider email lists; local housing authorities; Nichols Research, a California-based market research firm; and the Ipsos™ KnowledgePanel, an online random probability panel. See the documentation for more information about sampling and weighting. ## Survey Records, Data, and Documentation In total, 7,406 households completed 19,084 surveys representing 150,012 trips across 42,567 person-days.

32 ENERGY CONSERVATION, CONSUMPTION, AND UTILIZATI↗

2022 Central California Travel Study

# 2022 Central California Travel Study The 2022 Central California Travel Study collected demographic and travel pattern information to better understand household travel behavior in the state’s San Joaquin Valley region. ## Data Collection Agency The Fresno Council of Governments conducted the household travel study in collaboration with metropolitan planning organizations in Fresno, Kern, Kings, Madera, Merced, San Joaquin, Stanislaus, and Tulare counties in California. ## Survey Methodology The two-part study consisted of a survey and a travel diary. The survey gathered data on household demographic composition and typical travel behaviors. The travel diary gathered individual travel data during a specified travel period for all members of a given household. ## Travel Diary Methods Households with smartphones were encouraged to complete their travel diaries using the rMove smartphone app for up to seven consecutive days. Households without smartphones or those unwilling to participate via smartphone completed their travel diaries online (using the web-version of rMove) or by calling the survey call center. These households reported travel for one day (Tuesday, Wednesday, or Thursday). ## Recruitment Sampling Methods The majority of the recruitment was conducted via address-based sampling, a type of probability sampling, with a focus on reaching county-level targets in collaboration with metropolitan planning organizations in the region. Supplemental sampling methods, primarily non-probability, were employed during all waves of data collection to improve survey representation. The supplemental sample included targeted outreach to hard-to-survey populations via transit rider email lists; local housing authorities; Nichols Research, a California-based market research firm; and the Ipsos™ KnowledgePanel, an online random probability panel. See the documentation for more information about sampling and weighting. ## Survey Records, Data, and Documentation In total, 7,406 households completed 19,084 surveys representing 150,012 trips across 42,567 person-days.

32 ENERGY CONSERVATION, CONSUMPTION, AND UTILIZATI↗

2022 Central California Travel Study

# 2022 Central California Travel Study The 2022 Central California Travel Study collected demographic and travel pattern information to better understand household travel behavior in the state’s San Joaquin Valley region. ## Data Collection Agency The Fresno Council of Governments conducted the household travel study in collaboration with metropolitan planning organizations in Fresno, Kern, Kings, Madera, Merced, San Joaquin, Stanislaus, and Tulare counties in California. ## Survey Methodology The two-part study consisted of a survey and a travel diary. The survey gathered data on household demographic composition and typical travel behaviors. The travel diary gathered individual travel data during a specified travel period for all members of a given household. ## Travel Diary Methods Households with smartphones were encouraged to complete their travel diaries using the rMove smartphone app for up to seven consecutive days. Households without smartphones or those unwilling to participate via smartphone completed their travel diaries online (using the web-version of rMove) or by calling the survey call center. These households reported travel for one day (Tuesday, Wednesday, or Thursday). ## Recruitment Sampling Methods The majority of the recruitment was conducted via address-based sampling, a type of probability sampling, with a focus on reaching county-level targets in collaboration with metropolitan planning organizations in the region. Supplemental sampling methods, primarily non-probability, were employed during all waves of data collection to improve survey representation. The supplemental sample included targeted outreach to hard-to-survey populations via transit rider email lists; local housing authorities; Nichols Research, a California-based market research firm; and the Ipsos™ KnowledgePanel, an online random probability panel. See the documentation for more information about sampling and weighting. ## Survey Records, Data, and Documentation In total, 7,406 households completed 19,084 surveys representing 150,012 trips across 42,567 person-days.

32 ENERGY CONSERVATION, CONSUMPTION, AND UTILIZATI↗

2022 Central California Travel Study

# 2022 Central California Travel Study The 2022 Central California Travel Study collected demographic and travel pattern information to better understand household travel behavior in the state’s San Joaquin Valley region. ## Data Collection Agency The Fresno Council of Governments conducted the household travel study in collaboration with metropolitan planning organizations in Fresno, Kern, Kings, Madera, Merced, San Joaquin, Stanislaus, and Tulare counties in California. ## Survey Methodology The two-part study consisted of a survey and a travel diary. The survey gathered data on household demographic composition and typical travel behaviors. The travel diary gathered individual travel data during a specified travel period for all members of a given household. ## Travel Diary Methods Households with smartphones were encouraged to complete their travel diaries using the rMove smartphone app for up to seven consecutive days. Households without smartphones or those unwilling to participate via smartphone completed their travel diaries online (using the web-version of rMove) or by calling the survey call center. These households reported travel for one day (Tuesday, Wednesday, or Thursday). ## Recruitment Sampling Methods The majority of the recruitment was conducted via address-based sampling, a type of probability sampling, with a focus on reaching county-level targets in collaboration with metropolitan planning organizations in the region. Supplemental sampling methods, primarily non-probability, were employed during all waves of data collection to improve survey representation. The supplemental sample included targeted outreach to hard-to-survey populations via transit rider email lists; local housing authorities; Nichols Research, a California-based market research firm; and the Ipsos™ KnowledgePanel, an online random probability panel. See the documentation for more information about sampling and weighting. ## Survey Records, Data, and Documentation In total, 7,406 households completed 19,084 surveys representing 150,012 trips across 42,567 person-days.

32 ENERGY CONSERVATION, CONSUMPTION, AND UTILIZATI↗

2022 Central California Travel Study

# 2022 Central California Travel Study The 2022 Central California Travel Study collected demographic and travel pattern information to better understand household travel behavior in the state’s San Joaquin Valley region. ## Data Collection Agency The Fresno Council of Governments conducted the household travel study in collaboration with metropolitan planning organizations in Fresno, Kern, Kings, Madera, Merced, San Joaquin, Stanislaus, and Tulare counties in California. ## Survey Methodology The two-part study consisted of a survey and a travel diary. The survey gathered data on household demographic composition and typical travel behaviors. The travel diary gathered individual travel data during a specified travel period for all members of a given household. ## Travel Diary Methods Households with smartphones were encouraged to complete their travel diaries using the rMove smartphone app for up to seven consecutive days. Households without smartphones or those unwilling to participate via smartphone completed their travel diaries online (using the web-version of rMove) or by calling the survey call center. These households reported travel for one day (Tuesday, Wednesday, or Thursday). ## Recruitment Sampling Methods The majority of the recruitment was conducted via address-based sampling, a type of probability sampling, with a focus on reaching county-level targets in collaboration with metropolitan planning organizations in the region. Supplemental sampling methods, primarily non-probability, were employed during all waves of data collection to improve survey representation. The supplemental sample included targeted outreach to hard-to-survey populations via transit rider email lists; local housing authorities; Nichols Research, a California-based market research firm; and the Ipsos™ KnowledgePanel, an online random probability panel. See the documentation for more information about sampling and weighting. ## Survey Records, Data, and Documentation In total, 7,406 households completed 19,084 surveys representing 150,012 trips across 42,567 person-days.

32 ENERGY CONSERVATION, CONSUMPTION, AND UTILIZATI↗