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At least 19 records

Biochemical inhibition of acid phosphatase activity in two mountain spruce forest soils

The product inhibition of the 4-Methylumbelliferyl phosphate (MUB-P) decay rate, a measure of potential acid phosphatase activity, has not been considered in most of the published kinetic studies. The aim of this study was to determine the type and strength of the product inhibition in order to better define reaction conditions at which the acid phosphatase activity assay produces unbiased results. The MUB-P decay rate was measured in the forest floor and topsoil organic horizons of two spruce forest catchments at different initial MUB-P and P-PO4 3- concentrations. Furthermore, the type and strength of the inhibition was analyzed by non-linear regression. We found that MUB-P decay was competitively inhibited by the P-PO4 3- but also by dissolved organic P. By using estimated kinetic parameters, we calculated an underestimation of potential acid phosphatase activity by up to ~20% at concentrations of added MUB-P as high as 300 µmol g-1 dry weight and an incubation time one hour due to background inorganic and organic P concentrations in range of units of µmol per gram of dry soil. We discuss approaches that can be used to minimize the underestimation and show that the previously defined recommendations might not be always sufficient to avoid the bias. Therefore, we recommend to analyze progress curves at a wide range of initial MUB-P concentrations using non-linear methods when the enzyme assay is optimized for a given soil. When changes in inorganic and/or organic P concentrations are expected, we further recommend measuring background inorganic and organic P concentrations.

59 BASIC BIOLOGICAL SCIENCES↗

Bringing function to structure: Root–soil interactions shaping phosphatase activity throughout a soil profile in Puerto Rico

Abstract Large areas of highly productive tropical forests occur on weathered soils with low concentrations of available phosphorus (P). In such forests, root and microbial production of acid phosphatase enzymes capable of mineralizing organic phosphorus is considered vital to increasing available P for plant uptake. We measured both root and soil phosphatase throughout depth and alongside a variety of root and soil factors to better understand the potential of roots and soil biota to increase P availability and to constrain estimates of the biochemical mineralization within ecosystem models. We measured soil phosphatase down to 1 m, root phosphatase to 30 cm, and collected data on fine‐root mass density, specific root length, soil P, bulk density, and soil texture using soil cores in four tropical forests within the Luquillo Experimental Forest in Puerto Rico. We found that soil phosphatase decreased with soil depth, but not root phosphatase. Furthermore, when both soil and root phosphatase were expressed per soil volume, soil phosphatase was 100‐fold higher that root phosphatase. Both root and soil factors influenced soil and root phosphatase. Soil phosphatase increased with fine‐root mass density and organic P, which together explained over 50% of the variation in soil phosphatase. Over 80% of the variation in root phosphatase per unit root mass was attributed to specific root length (positive correlation) and available (resin) P (negative correlation). Synthesis : Fine‐root traits and soil P data are necessary to understand and represent soil and root phosphatase activity throughout the soil column and across sites with different soil conditions and tree species. These findings can be used to parameterize or benchmark estimates of biochemical mineralization in ecosystem models that contain fine‐root biomass and soil P distributions throughout depth.

59 BASIC BIOLOGICAL SCIENCES↗

Low soil phosphorus availability triggers maize growth stage specific rhizosphere processes leading to mineralization of organic P

Phosphorus (P) is one of the essential nutrients for all living organisms. High-quality mineral P fertilizer is a finite resource with only an estimated 80-100 years of reserves remaining globally. Both plants and microbes have developed several mechanisms such as secretion of organic acids, acidification of the rhizosphere, and production of extracellular enzymes to enhance the acquisition of P from soils. Increased activity of phosphatase occurs in response to P deficiency as part of P starvation responses which catalyze the hydrolysis of P o from soil organic matter (SOM) and release inorganic P (P i ). However, the specific interactions between plants and microbial communities in the rhizosphere soil triggered by P deficiency are largely unknown. In a greenhouse study we grew maize plants in low and high P containing soils and examined the role of rhizosphere processes on mineralization of organic P (P o ) in soils with low and high P availability and different P speciation 23 (Po dominated versus P i dominated) and determine its effect on plant growth. Plant biomass was determined and rhizosphere soils, and soil samples from pots with and without plants were collected at key vegetative growth stages (VGS). A variety of biogeochemical parameters (e.g. microbial biomass C and P, potential phosphatase activities, plant biomass P, soil P species etc.) were determined using both conventional (sequential chemical extraction) and advanced (e.g. high resolution mass spectrometry (FTICR-MS) and 3129 P solution NMR) techniques. Here, we used phospholipid fatty acid (PLFA) and DNA based microbial community analysis to track changes in microbial community structure and diversity. Low P availability induced changes in biogeochemical processes and microbial community composition in the rhizosphere soils of plants grown in low P containing soil. At early VGS, the plants in low P soil were visually struggling which correlated with greater rhizosphere potential acid phosphatase activity and a reduction in P o fractions. At late VGS, the plants appeared to recover which correlated with a decrease in Meh (III) extractable P, an increase in microbial biomass C and P and greater total P in the plant biomass (roots, shoots and leaves). In high P containing soil, on the other hand, greater P availability masked these rhizosphere processes. Our results confirmed the degradation of SOM and Po mineralization in rhizosphere soils driven by microbe and plant need for P. P deficiency may have favored the abundance of fungi which utilized easily degradable root exudates for the production of extracellular enzymes responsible for the mineralize of SOM P o . This study highlights the intertwined mechanisms involved, and emphasizes the importance of, P o sources for plant nutrition and survival in marginal soils.

59 BASIC BIOLOGICAL SCIENCES↗

Trade-Offs in Phosphorus Acquisition Strategies of Five Common Tree Species in a Tropical Forest of Puerto Rico

Tree species that are successful in tropical lowlands have different acquisition strategies to overcome soil phosphorus (P) limitations. Some of these strategies belowground include adjustments in fine-root traits, such as morphology, architecture, association with arbuscular mycorrhizal fungi, and phosphatase activity. Trade-offs among P-acquisition strategies are expected because of their respective carbon cost. However, empirical evidence remains scarce which hinders our understanding of soil P-acquisition processes in tropical forests. Here, we measured seven fine-root functional traits related to P acquisition of five common tree species in three sites of the Luquillo Experimental Forest in Puerto Rico. We then described species-specific P-acquisition strategies and explored the changes in fine-root trait expression from 6 months before to 6 months after two consecutive hurricanes, Irma and María, passed over the island. We found that variations in root trait expression were driven mainly by the large interspecific differences across the three selected sites. In addition, we revealed a trade-off between highly colonized fine roots with high phosphatase activity and fine roots that have a high degree of branching. Furthermore, the former strategy was adopted by pioneer species ( Spathodea campanulata and Cecropia schreberiana ), whereas the latter was adopted by non-pioneer species (mostly Dacryodes excelsa and Prestoea montana ). Additionally, we found that root trait expression did not change comparing 6 months before and after the hurricanes, with the exception of root phosphatase activity. Altogether, our results suggest a combination of structural and physiological root traits for soil P acquisition in P-poor tropical soils by common tropical tree species, and show stability on most of the root trait expression after hurricane disturbances.

59 BASIC BIOLOGICAL SCIENCES↗

Effects of experimental and seasonal drying on soil microbial biomass and nutrient cycling in four lowland tropical forests

Changes in precipitation represent a major effect of climate change on tropical forests, which contain some of the earth’s largest terrestrial carbon (C) stocks. Such changes are expected to influence microbes, nutrients, and the fate of C in tropical forest soils. To explore this, we assessed soil microbial biomass, potential extracellular enzyme activities, and nutrient availability in a partial throughfall exclusion experiment in four seasonal lowland tropical humid forests in Panama with wide variation in precipitation and soil fertility. We hypothesized that throughfall exclusion would reduce microbial biomass and activity and accentuate dry season soil nutrient accumulation, with larger effects in wetter, less droughtresistant forests. We observed a baseline seasonal pattern of decreased microbial biomass and increased extractable dissolved organic C (DOC), total dissolved nitrogen (TDN), nitrate (NO 3 – ), and resinextractable phosphorus (P) in the dry season, with the strongest patterns for nitrogen (N). However, potential enzyme activities showed no consistent seasonality. In line with seasonal drying, throughfall exclusion decreased soil microbial biomass in the wet season and increased TDN and NO 3 – , especially in the dry season. In contrast to seasonal drying, throughfall exclusion decreased DOC and did not affect resinextractable P, but slightly decreased potential phosphatase activities. Potential enzyme activities varied among sites and sampling times, but did not explain much variation in microbial biomass or substrate availability. We conclude that reduced rainfall in tropical forests might accentuate some dry season patterns, like reductions in microbial biomass and accumulation of extractable nutrients. However, our data also suggest new patterns, like reduced inputs of DOC to soils with drying, which could have cascading effects on soil ecological function and C storage.

54 ENVIRONMENTAL SCIENCES↗

High-Throughput Microfluidics Platform for Intracellular Delivery and Sampling of Biomolecules from Live Cells

Nondestructive cell membrane permeabilization systems enable the intracellular delivery of exogenous biomolecules for cell engineering tasks as well as the temporal sampling of cytosolic contents from live cells for the analysis of dynamic processes. Here, in this paper, we report a microwell array format live-cell analysis device (LCAD) that can perform localized-electroporation induced membrane permeabilization, for cellular delivery or sampling, and directly interfaces with surface-based biosensors for analyzing the extracted contents. We demonstrate the capabilities of the LCAD via an automated high-throughput workflow for multimodal analysis of live-cell dynamics, consisting of quantitative measurements of enzyme activity using self-assembled monolayers for MALDI mass spectrometry (SAMDI) and deep-learning enhanced imaging and analysis. By combining a fabrication protocol that enables robust assembly and operation of multilayer devices with embedded gold electrodes and an automated imaging workflow, we successfully deliver functional molecules (plasmid and siRNA) into live cells at multiple time-points and track their effect on gene expression and cell morphology temporally. Furthermore, we report sampling performance enhancements, achieving saturation levels of protein tyrosine phosphatase activity measured from as few as 60 cells, and demonstrate control over the amount of sampled contents by optimization of electroporation parameters using a lumped model. Lastly, we investigate the implications of cell morphology on electroporation-induced sampling of fluorescent molecules using a deep-learning enhanced image analysis workflow.

59 BASIC BIOLOGICAL SCIENCES↗

Sac1 links phosphoinositide turnover to cryptococcal virulence

Cryptococcus neoformans is an environmentally acquired fungal pathogen that causes over 140,000 deaths per year. Cryptococcal infection occurs when infectious particles are deposited into the lung, where they encounter host phagocytic cells. C. neoformans may be engulfed by these phagocytes, an important step of infection that leads to outcomes ranging from termination of infection to cryptococcal dissemination. To study this critical process, we screened approximately 4,700 cryptococcal gene deletion mutants for altered uptake, using primary mouse and human phagocytic cells. Among the hits of these two screens, we identified 93 mutants with perturbed uptake in both systems, as well as others with differences in uptake by only one cell type. We further screened the hits for changes in thickness of the capsule, a protective polysaccharide layer around the cell which is an important cryptococcal virulence factor. The combination of our three screens yielded 45 mutants, including one lacking the phosphatidylinositol-4-phosphate phosphatase Sac1. In this work, we implicate Sac1 in both host cell uptake and capsule production. We found that sac1 mutants exhibit lipid trafficking defects, reductions in secretory system function, and changes in capsule size and composition. Many of these changes occur specifically in tissue culture media, highlighting the role of Sac1 phosphatase activity in responding to the stress of host-like conditions. Overall, these findings show how genome-scale screening can identify cellular factors that contribute to our understanding of cryptococcal biology and demonstrate the role of Sac1 in determining fungal virulence.

59 BASIC BIOLOGICAL SCIENCES↗

B56$δ$ long-disordered arms form a dynamic PP2A regulation interface coupled with global allostery and Jordan’s syndrome mutations

Intrinsically disordered regions (IDR) and short linear motifs (SLiMs) play pivotal roles in the intricate signaling networks governed by phosphatases and kinases. B56δ (encoded by PPP2R5D) is a regulatory subunit of protein phosphatase 2A (PP2A) with long IDRs that harbor a substrate-mimicking SLiM and multiple phosphorylation sites. De novo missense mutations in PPP2R5D cause intellectual disabilities (ID), macrocephaly, Parkinsonism, and a broad range of neurological symptoms. Our single-particle cryo-EM structures of the PP2A-B56δ holoenzyme reveal that the long, disordered arms at the B56δ termini fold against each other and the holoenzyme core. This architecture suppresses both the phosphatase active site and the substrate-binding protein groove, thereby stabilizing the enzyme in a closed latent form with dual autoinhibition. The resulting interface spans over 190 Å and harbors unfavorable contacts, activation phosphorylation sites, and nearly all residues with ID-associated mutations. Our studies suggest that this dynamic interface is coupled to an allosteric network responsive to phosphorylation and altered globally by mutations. Furthermore, we found that ID mutations increase the holoenzyme activity and perturb the phosphorylation rates, and the severe variants significantly increase the mitotic duration and error rates compared to the normal variant.

59 BASIC BIOLOGICAL SCIENCES↗

Phosphorus limitation of early growth differs between nitrogen‐fixing and nonfixing dry tropical forest tree species

Summary Tropical forests are often characterized by low soil phosphorus (P) availability, suggesting that P limits plant performance. However, how seedlings from different functional types respond to soil P availability is poorly known but important for understanding and modeling forest dynamics under changing environmental conditions. We grew four nitrogen (N)‐fixing Fabaceae and seven diverse non‐N‐fixing tropical dry forest tree species in a shade house under three P fertilization treatments and evaluated carbon (C) allocation responses, P demand, P‐use, investment in P acquisition traits, and correlations among P acquisition traits. Nitrogen fixers grew larger with increasing P addition in contrast to non‐N fixers, which showed fewer responses in C allocation and P use. Foliar P increased with P addition for both functional types, while P acquisition strategies did not vary among treatments but differed between functional types, with N fixers showing higher root phosphatase activity (RPA) than nonfixers. Growth responses suggest that N fixers are limited by P, but nonfixers may be limited by other resources. However, regardless of limitation, P acquisition traits such as mycorrhizal colonization and RPA were nonplastic across a steep P gradient. Differential limitation among plant functional types has implications for forest succession and earth system models.

59 BASIC BIOLOGICAL SCIENCES↗

PTEN protects kidney against acute kidney injury by alleviating apoptosis and promoting autophagy via regulating HIF1-α and mTOR through PI3K/Akt pathway

Highlights: • PTEN protects kidney against acute kidney injury. • The effect of PTEN works by alleviating apoptosis and promoting autophagy via regulating HIF1-α and mTOR by PI3K/Akt pathway. Phosphatase and tensin homolog (PTEN) deleted on human chromosome 10 is a tumor suppressor with bispecific phosphatase activity, which is often involved in the study of energy metabolism and tumorigenesis. PTEN is recently reported to participate in the process of acute injury. However, the mechanism of PTEN in Ischemia-Reperfusion Injury (IRI) has not yet been clearly elucidated. In this study, mice with bilateral renal artery ischemia-reperfusion and HK-2 cells with hypoxia/reoxygenation (H/R) were used as acute kidney injury models. We demonstrated that PTEN was downregulated in IRI-induced kidney as well as in H/R-induced HK-2 cells. By silencing and overexpressing PTEN with si-PTEN RNA and PHBLV-CMV-PTEN-flag lentivirus before H/R, we found that PTEN protected HK-2 cells against H/R-induced injury reflected by the change in cell activity and the release of LDH. Furthermore, we inhibited HIF1-α with PX-478 and inactivated mTOR with Rapamycin before the silence of PTEN in H/R model. Our data indicated that the renoprotective effect of PTEN worked via PI3K/Akt/mTOR pathway and PI3K/Akt/HIF1-α pathway, hence alleviating apoptosis and improving autophagy respectively. Our findings provide valuable insights into the molecular mechanism underlying renoprotection of PTEN on autophagy and apoptosis induced by renal IRI, which offers a novel therapeutic target for the treatment of AKI.

60 APPLIED LIFE SCIENCES↗

Injectable platelet rich fibrin facilitates hair follicle regeneration by promoting human dermal papilla cell proliferation, migration, and trichogenic inductivity

Hair follicle regeneration has been successful in mice but failed in human being for years. Dermal papilla cells, a specialized mesenchymal stem cell derived from dermal papilla within hair follicles, is considered the key cells for hair follicle regeneration function as both regeneration initiator and regulator. Injectable platelet rich fibrin (i-PRF), a novel biomaterial rich in a variety of growth factors and three-dimensional scaffolds, has shown promising effects on tissue regeneration. In this study, we aimed to evaluate the application of i-PRF in human hair follicle regeneration by examining the biological effects of i-PRF on human dermal papilla cells (hDPCs). Biomaterial compatibility, cell viability, proliferation, migration, alkaline phosphatase activity and trichogenic inductivity were assessed after exposing hDPCs to different concentrations of i-PRF extracts. In addition, we investigated the ultrastructure of i-PRF with all cell components filtered. The results revealed that i-PRF possessing excellent biocompatibility and could significantly promote hDPCs proliferation, migration, and trichogenic inductivity. Furthermore, the concentration of i-PRF is able to remarkably influence hDPCs behavior in a dose-dependent pattern. Different concentrations exhibited differential effects on hDPCs behavior. In general, lower concentration promotes cell proliferation better than higher concentration, while higher concentration promotes cell function better reversely. Best concentration for hDPCs in vitro expending is 1% concentration. 20% concentration is optimal for hair follicle regeneration. In summary, our findings concluded that i-PRF facilitates hair follicle regeneration by promoting human dermal papilla cell proliferation, migration, and trichogenic inductivity.

60 APPLIED LIFE SCIENCES↗

Plant responses to elevated CO 2 under competing hypotheses of nitrogen and phosphorus limitations

Abstract The future ecosystem carbon cycle has important implications for biosphere‐climate feedback. The magnitude of future plant growth and carbon accumulation depends on plant strategies for nutrient uptake under the stresses of nitrogen (N) versus phosphorus (P) limitations. Two archetypal theories have been widely acknowledged in the literature to represent N and P limitations on ecosystem processes: Liebig's Law of the Minimum (LLM) and the Multiple Element Limitation (MEL) approach. LLM states that the more limiting nutrient controls plant growth, and commonly leads to predictions of dramatically dampened ecosystem carbon accumulation over the 21st century. Conversely, the MEL approach recognizes that plants possess multiple pathways to coordinate N and P availability and invest resources to alleviate N or P limitation. We implemented these two contrasting approaches in the E3SM model, and compiled 98 in situ forest N or P fertilization experiments to evaluate how terrestrial ecosystems will respond to N and P limitations. We find that MEL better captured the observed plant responses to nutrient perturbations globally, compared with LLM. Furthermore, LLM and MEL diverged dramatically in responses to elevated CO 2 concentrations, leading to a two‐fold difference in CO 2 fertilization effects on Net Primary Productivity by the end of the 21st century. The larger CO 2 fertilization effects indicated by MEL mainly resulted from plant mediation on N and P resource supplies through N 2 fixation and phosphatase activities. This analysis provides quantitative evidence of how different N and P limitation strategies can diversely affect future carbon and nutrient dynamics.

54 ENVIRONMENTAL SCIENCES↗

Root exudates induced coupled carbon and phosphorus cycling in a soil with low phosphorus availability

The amount and type of root exudates can influence P availability in the rhizosphere directly by desorption or dissolution of soil minerals, or indirectly by decomposition of soil organic matter (SOM). Here this study aimed to determine the mechanisms by which specific root exudates influence the distribution and availability of P in soils with low P availability. Water, glucose, alanine, and oxalate were delivered through a simulated root into soils for 15 days. Zymography and planar optodes were used to image potential phosphatase activity, and O2 and pH distribution, respectively. Soils were analyzed for resin extractable inorganic P (Pi), dissolved organic C (DOC), water soluble Fe, and Al, and microbial community structure. Characterization of SOM and P were conducted using ultra-high resolution mass spectrometry and 31P solution nuclear magnetic resonance (NMR), respectively. The addition of oxalate resulted in the greatest resin extractable Pi, DOC, and water-soluble Fe, and Al compared to the other exudates suggesting destabilization of mineral associated organic matter (MAOM) and release of organic P (Po). Both 31P solution NMR and ultra-high resolution mass spectrometry analysis provided evidence of mineralization of Po released from the destabilization of MAOM. The study demonstrates the important role microbial and plant-derived metal chelating ligands play in destabilizing MAOM, releasing SOM and importantly Po, that when mineralized may contribute to increasing Pi availability in soils with low P availability.

59 BASIC BIOLOGICAL SCIENCES↗

Influence of Uranium Concentration and pH on U-Phosphate Biomineralization by Caulobacter OR37

Uranium contamination of soils and groundwater in the United States represents a significant health risk and will require multiple remediation approaches. Microbial phosphatase activity coupled to the addition of an organic P source has recently been studied as a remediation strategy that provides an extended release of inorganic P (Pi) into U-contaminated sites, resulting in the precipitation of meta-autunite minerals. Previous laboratory- and field-based biomineralization studies have investigated environments with relatively high U concentrations (>20 μM). However, most contaminated sites have much lower U concentrations (<2 μM). The Environmental Protection Agency (EPA) limit for U in drinking water is 0.126 μM. Reaching this regulatory limit becomes challenging as U concentrations approach autunite solubility. Furthermore, we studied the precipitation of U(VI)-phosphate minerals by an environmental isolate of Caulobacter sp. (strain OR37) from an Oak Ridge, Tennessee, U-contaminated site. Abiotic U(VI) solubility experiments reveal that U(VI)-phosphate minerals do not form in the presence of excess Pi (500 μM) when U(VI) concentrations are <1 μM and pH is <5. When OR37 cells are reacted under the same conditions with Pi or glycerol-2-phosphate, U(VI)-phosphate mineral formation was observed, along with the formation of intracellular polyphosphate granules. These results show that bacteria provide supersaturated microenvironments needed for U(VI)-phosphate mineralization while hydrolyzing organic P sources. This provides a pathway to lower U concentrations to below EPA limits for drinking water.

38 RADIATION CHEMISTRY, RADIOCHEMISTRY, AND NUCLEA↗

Rooting for function: community‐level fine‐root traits relate to many ecosystem functions

Humans are driving biodiversity change, which also alters community functional traits. However, how changes in the functional traits of the community alter ecosystem functions—especially belowground—remains an important gap in our understanding of the consequences of biodiversity change. We test hypotheses for how the root traits of the root economics space (composed of the collaboration and conservation gradients) are associated with proxies for ecosystem functioning across grassland and forest ecosystems in both observational and experimental datasets from 810 plant communities. First, we assessed whether community-weighted means of the root economics space traits adhered to the same trade-offs as species-level root traits. Then, we examined the relationships between community-weighted mean root traits and aboveground biomass production, root standing biomass, soil fauna biomass, soil microbial biomass, decomposition of standard and plot-specific material, ammonification, nitrification, phosphatase activity, and drought resistance. We found evidence for a community collaboration gradient but not for a community conservation gradient. Yet, links between community root traits and ecosystem functions were more common than we expected, especially for aboveground biomass, microbial biomass, and decomposition. These findings suggest that changes in species composition, which alter root trait means, will in turn affect critical ecosystem functions.

54 ENVIRONMENTAL SCIENCES↗

Fine-root traits from common tree species in Puerto Rico before and after Hurricane María (2017-2018)

This dataset is a compilation of root trait measurements from common tree species in the Luquillo Experimental Forest in Puerto Rico taken in February 2017 (before Hurricane María), and in February 2018 (after the hurricane). This dataset is being used on a publication called "Tradeoffs in phosphorus acquisition strategies of five tree species in a tropical forest of Puerto Rico", where we describe belowground P-acquisition strategies of five common tree species (two pioneers and three non-pioneers) growing in the lowland tropical wet forests of Puerto Rico. Additionally, we used the unique opportunity of two consecutive hurricanes that passed over the island (Irma and Maria) to compare root traits before and after the hurricanes to determine which traits were the most responsive and whether such disturbance affected root trait relationships and trade-offs. For this, we sampled trees from EV1, SB2, and NGEE Tropics' plots EVV and EVR in 2017 (before hurricanes) and 2018 (after hurricanes). Morphological fine-root traits include root length, diameter, and SRL. Root architectural traits includes root branching ratio and intensity. Root physiological trait measured includes phosphatase activity, and root microbial association trait measured includes percentage mycorrhizal colonization. Chemistry data includes root phosphorus concentration, soil available, and organic phosphorus.

54 ENVIRONMENTAL SCIENCES↗

Godzilla mineral dust and La Soufrière volcanic ash fallout immediately stimulate marine microbial phosphate uptake

During the “Godzilla” dust storm of June 2020, unusually high fluxes of mineral dust traveled across the Atlantic from the Sahara Desert, reaching the Caribbean Basin, Gulf Coast, and southeastern United States. Additionally, an eruption of the La Soufrière volcano on St. Vincent in April 2021 generated substantial ashfall in the southeastern Caribbean. While many studies have analyzed mineral dust’s ability to relieve nutrient limitation of phosphorus (P) in the P-stressed North Atlantic, less is known about the impact of extreme events and other natural aerosols on fluxes of P into seawater and from seawater into marine microbial cells. We quantified P and iron (Fe) content in mineral dust from the Godzilla dust storm and volcanic ash from the La Soufrière eruption collected at Ragged Point, Barbados. We also performed seawater incubations to assess the marine microbial response to aerosol deposition. Using environmentally-relevant concentrations of atmospheric particles for within the ocean’s mixed layer allowed us to draw realistic conclusions about how these deposition events impacted P cycling in situ. Volcanic ash has lower P content than mineral dust, and P in volcanic ash is far less soluble (~1%) than assumed in current atmospheric deposition models. Adding mineral dust and the volcanic ash leachate in concentrations representing different deposition scenarios increased soluble reactive phosphorus (SRP) concentrations in coastal seawater by ~7-32 nM. Phosphate uptake rate was stimulated in coastal seawater after either mineral dust or volcanic ash deposition at aerosol concentrations relevant to the Godzilla dust event, with ash eliciting the fastest uptake rate. Furthermore, high concentrations of both the mineral dust and volcanic ash led to slightly elevated alkaline phosphatase activity (APA) compared to the relevant controls, indicating higher potential for use of dissolved organic phosphorus (DOP) as a P source. Quantifying these aerosols’ impacts on P cycling is a significant step towards achieving a better understanding of their potential roles in relieving nutrient limitation and fueling the biological carbon pump.

54 ENVIRONMENTAL SCIENCES↗

Dissecting protein tyrosine phosphatase signaling by engineered chemogenetic control of its activity

Protein tyrosine phosphatases (PTPases) are critical mediators of dynamic cell signaling. A tool capable of identifying transient signaling events downstream of PTPases is essential to understand phosphatase function on a physiological time scale. We report a broadly applicable protein engineering method for allosteric regulation of PTPases. This method enables dissection of transient events and reconstruction of individual signaling pathways. Implementation of this approach for Shp2 phosphatase revealed parallel MAPK and ROCK II dependent pathways downstream of Shp2, mediating transient cell spreading and migration. Furthermore, we show that the N-SH2 domain of Shp2 regulates MAPK-independent, ROCK II-dependent cell migration. Engineered targeting of Shp2 activity to different protein complexes revealed that Shp2-FAK signaling induces cell spreading whereas Shp2-Gab1 or Shp2-Gab2 mediates cell migration. We identified specific transient morphodynamic processes induced by Shp2 and determined the role of individual signaling pathways downstream of Shp2 in regulating these events. Broad application of this approach is demonstrated by regulating PTP1B and PTP-PEST phosphatases.

59 BASIC BIOLOGICAL SCIENCES↗