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Fanzor (Fz) is an ωRNA-guided endonuclease extensively found throughout the eukaryotic domain with unique gene editing potential. Here, we describe the structures of Fzs from three different organisms. We find that Fzs share a common ωRNA interaction interface, regardless of the length of the ωRNA, which varies considerably across species. The analysis also reveals Fz's mode of DNA recognition and unwinding capabilities as well as the presence of a non-canonical catalytic site. The structures demonstrate how protein conformations of Fz shift to allow the binding of double-stranded DNA to the active site within the R-loop. Mechanistically, examination of structures in different states shows that the conformation of the lid loop on the RuvC domain is controlled by the formation of the guide/DNA heteroduplex, regulating the activation of nuclease and DNA double-stranded displacement at the single cleavage site. Our findings clarify the mechanism of Fz, establishing a foundation for engineering efforts.
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The last steps in mRNA export and remodeling are performed by the Nup82 complex, a large conserved assembly at the cytoplasmic face of the nuclear pore complex (NPC). By integrating diverse structural data, we have determined the molecular architecture of the native Nup82 complex at subnanometer precision. The complex consists of two compositionally identical multiprotein subunits that adopt different configurations. The Nup82 complex fits into the NPC through the outer ring Nup84 complex. Our map shows that this entire 14-MDa Nup82-Nup84 complex assembly positions the cytoplasmic mRNA export factor docking sites and messenger ribonucleoprotein (mRNP) remodeling machinery right over the NPC's central channel rather than on distal cytoplasmic filaments, as previously supposed. We suggest that this configuration efficiently captures and remodels exporting mRNP particles immediately upon reaching the cytoplasmic side of the NPC.
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Proteínas de Complejo Poro Nuclear/química , Poro Nuclear/química , Proteínas de Saccharomyces cerevisiae/metabolismo , Saccharomyces cerevisiae/metabolismo , Levaduras/metabolismo , Transporte Activo de Núcleo Celular , Proteínas Fúngicas , Proteínas de Complejo Poro Nuclear/ultraestructura , ARN Mensajero , Saccharomyces cerevisiae/citología , Proteínas de Saccharomyces cerevisiae/ultraestructuraRESUMEN
Citrate is best known as an intermediate in the tricarboxylic acid cycle of the cell. In addition to this essential role in energy metabolism, the tricarboxylate anion also acts as both a precursor and a regulator of fatty acid synthesis1-3. Thus, the rate of fatty acid synthesis correlates directly with the cytosolic concentration of citrate4,5. Liver cells import citrate through the sodium-dependent citrate transporter NaCT (encoded by SLC13A5) and, as a consequence, this protein is a potential target for anti-obesity drugs. Here, to understand the structural basis of its inhibition mechanism, we determined cryo-electron microscopy structures of human NaCT in complexes with citrate or a small-molecule inhibitor. These structures reveal how the inhibitor-which binds to the same site as citrate-arrests the transport cycle of NaCT. The NaCT-inhibitor structure also explains why the compound selectively inhibits NaCT over two homologous human dicarboxylate transporters, and suggests ways to further improve the affinity and selectivity. Finally, the NaCT structures provide a framework for understanding how various mutations abolish the transport activity of NaCT in the brain and thereby cause epilepsy associated with mutations in SLC13A5 in newborns (which is known as SLC13A5-epilepsy)6-8.
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Proteínas Portadoras/antagonistas & inhibidores , Proteínas Portadoras/química , Ácido Cítrico/metabolismo , Microscopía por Crioelectrón , Malatos/farmacología , Fenilbutiratos/farmacología , Simportadores/antagonistas & inhibidores , Simportadores/química , Sitios de Unión , Encéfalo/metabolismo , Proteínas Portadoras/genética , Proteínas Portadoras/ultraestructura , Ácido Cítrico/química , Transportadores de Ácidos Dicarboxílicos/química , Transportadores de Ácidos Dicarboxílicos/metabolismo , Epilepsia/genética , Epilepsia/metabolismo , Humanos , Malatos/química , Modelos Moleculares , Mutación , Fenilbutiratos/química , Multimerización de Proteína , Sodio/metabolismo , Especificidad por Sustrato/efectos de los fármacos , Especificidad por Sustrato/genética , Simportadores/genética , Simportadores/ultraestructuraRESUMEN
The human GABAB receptor-a member of the class C family of G-protein-coupled receptors (GPCRs)-mediates inhibitory neurotransmission and has been implicated in epilepsy, pain and addiction1. A unique GPCR that is known to require heterodimerization for function2-6, the GABAB receptor has two subunits, GABAB1 and GABAB2, that are structurally homologous but perform distinct and complementary functions. GABAB1 recognizes orthosteric ligands7,8, while GABAB2 couples with G proteins9-14. Each subunit is characterized by an extracellular Venus flytrap (VFT) module, a descending peptide linker, a seven-helix transmembrane domain and a cytoplasmic tail15. Although the VFT heterodimer structure has been resolved16, the structure of the full-length receptor and its transmembrane signalling mechanism remain unknown. Here we present a near full-length structure of the GABAB receptor, captured in an inactive state by cryo-electron microscopy. Our structure reveals several ligands that preassociate with the receptor, including two large endogenous phospholipids that are embedded within the transmembrane domains to maintain receptor integrity and modulate receptor function. We also identify a previously unknown heterodimer interface between transmembrane helices 3 and 5 of both subunits, which serves as a signature of the inactive conformation. A unique 'intersubunit latch' within this transmembrane interface maintains the inactive state, and its disruption leads to constitutive receptor activity.
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Microscopía por Crioelectrón , Receptores de GABA-B/química , Receptores de GABA-B/ultraestructura , Calcio/metabolismo , Etanolaminas/química , Etanolaminas/metabolismo , Humanos , Ligandos , Modelos Moleculares , Fosforilcolina/química , Fosforilcolina/metabolismo , Dominios Proteicos , Multimerización de Proteína , Subunidades de Proteína/química , Subunidades de Proteína/metabolismo , Receptores de GABA-B/metabolismo , Relación Estructura-ActividadRESUMEN
An amendment to this paper has been published and can be accessed via a link at the top of the paper.
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Extracellular vesicles of endosomal origin, exosomes, mediate intercellular communication by transporting substrates with a variety of functions related to tissue homeostasis and disease. Their diagnostic and therapeutic potential has been recognized for diseases such as cancer in which signaling defects are prominent. However, it is unclear to what extent exosomes and their cargo inform the progression of infectious diseases. We recently defined a subset of exosomes termed defensosomes that are mobilized during bacterial infection in a manner dependent on autophagy proteins. Through incorporating protein receptors on their surface, defensosomes mediated host defense by binding and inhibiting pore-forming toxins secreted by bacterial pathogens. Given this capacity to serve as decoys that interfere with surface protein interactions, we investigated the role of defensosomes during infection by Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2), the etiological agent of Coronavirus Disease 2019 (COVID-19). Consistent with a protective function, exosomes containing high levels of the viral receptor ACE2 in bronchoalveolar lavage fluid (BALF) from critically ill COVID-19 patients was associated with reduced intensive care unit (ICU) and hospitalization times. We found ACE2+ exosomes were induced by SARS-CoV-2 infection and activation of viral sensors in cell culture, which required the autophagy protein ATG16L1, defining these as defensosomes. We further demonstrate that ACE2+ defensosomes directly bind and block viral entry. These findings suggest that defensosomes may contribute to the antiviral response against SARS-CoV-2 and expand our knowledge on the regulation and effects of extracellular vesicles during infection.
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Enzima Convertidora de Angiotensina 2/metabolismo , COVID-19 , Humanos , Peptidil-Dipeptidasa A/metabolismo , Receptores Virales , SARS-CoV-2RESUMEN
INTRODUCTION: Surgical stabilization of rib fractures (SSRF) is associated with lower rates of mortality and fewer complications. This study evaluates whether the decision to undergo SSRF is associated with age, race, ethnicity, and insurance status and assesses associated clinical outcomes. METHODS: This retrospective analysis included patients ≥45 y old with rib fractures who underwent SSRF in the Trauma Quality Improvement Program from 2016 to 2020. Race, ethnicity, and insurance statuses were collected. Age in years was dichotomized into two groups: 45-64 and 65+. Outcomes included ventilator-associated pneumonia, unplanned endotracheal intubation, acute respiratory distress syndrome, in-hospital mortality, failure to rescue (FTR) after major complications, and FTR after respiratory complications. Logistic regression models were fit to evaluate outcomes, controlling for gender, body mass index, Injury Severity Score, flail chest, chronic obstructive pulmonary disease, congestive heart failure, and smoking. RESULTS: Two thousand eight hundred thirty-nine patients aged 45-64 and 1828 patients aged 65+ underwent SSRF. No significant difference in clinical outcomes was noted between these groups. Analysis showed that the association of SSRF with ventilator-associated pneumonia, unplanned intubation, acute respiratory distress syndrome, in-hospital mortality, FTR after a major complication, or FTR after a respiratory complication did not vary by age (P > 0.05). Black (odds ratio [OR] 0.67; 95% confidence interval [CI]: 0.59-0.77; P < 0.001), Hispanic (OR 0.80; 95% CI: 0.71-0.91; P < 0.001), and Medicaid (OR = 0.85; 95% CI = 0.76-0.95; P = 0.005) patients were less likely to receive SSRF. CONCLUSIONS: No differences in clinical outcomes were measured between adults aged 45-64 and ≥65 who underwent SSRF. Older age should not preclude patients from receiving SSRF. Further work is needed to improve underutilization in Black, Hispanic and Medicaid patients.
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In Figs. 2b and 3d of this Letter, the labels 'Dynamin 1' and 'Overlay' were inadvertently swapped. This has been corrected online.
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Membrane fission is a fundamental process in the regulation and remodelling of cell membranes. Dynamin, a large GTPase, mediates membrane fission by assembling around, constricting and cleaving the necks of budding vesicles1. Here we report a 3.75 Å resolution cryo-electron microscopy structure of the membrane-associated helical polymer of human dynamin-1 in the GMPPCP-bound state. The structure defines the helical symmetry of the dynamin polymer and the positions of its oligomeric interfaces, which were validated by cell-based endocytosis assays. Compared to the lipid-free tetramer form2, membrane-associated dynamin binds to the lipid bilayer with its pleckstrin homology domain (PHD) and self-assembles across the helical rungs via its guanine nucleotide-binding (GTPase) domain3. Notably, interaction with the membrane and helical assembly are accommodated by a severely bent bundle signalling element (BSE), which connects the GTPase domain to the rest of the protein. The BSE conformation is asymmetric across the inter-rung GTPase interface, and is unique compared to all known nucleotide-bound states of dynamin. The structure suggests that the BSE bends as a result of forces generated from the GTPase dimer interaction that are transferred across the stalk to the PHD and lipid membrane. Mutations that disrupted the BSE kink impaired endocytosis. We also report a 10.1 Å resolution cryo-electron microscopy map of a super-constricted dynamin polymer showing localized conformational changes at the BSE and GTPase domains, induced by GTP hydrolysis, that drive membrane constriction. Together, our results provide a structural basis for the mechanism of action of dynamin on the lipid membrane.
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Biopolímeros/química , Biopolímeros/metabolismo , Membrana Celular/metabolismo , Membrana Celular/ultraestructura , Microscopía por Crioelectrón , Dinamina I/metabolismo , Dinamina I/ultraestructura , Biopolímeros/genética , Membrana Celular/química , Dinamina I/química , Dinamina I/genética , Endocitosis/genética , Guanosina Trifosfato/análogos & derivados , Guanosina Trifosfato/química , Guanosina Trifosfato/metabolismo , Humanos , Hidrólisis , Modelos Moleculares , Proteínas Mutantes/química , Proteínas Mutantes/genética , Proteínas Mutantes/metabolismo , Proteínas Mutantes/ultraestructura , Mutación , Dominios Proteicos , Multimerización de ProteínaRESUMEN
A significant cost of wildfires is the exposure of local and regional populations to air pollution from smoke, which can travel hundreds of miles from the source fire and is associated with significant negative health consequences. Wildfires are increasing in frequency and intensity in the United States, driven by historic fire management approaches and global climate change. These influences will take many decades or longer to reverse, so the main opportunities for mitigating health effects involve minimizing human exposure through changes in behavior or infrastructure. One key recommendation for reducing pollution exposures during wildfire smoke events is to limit time and physical activity outdoors, but there is limited evidence on the extent to which people make this change. We estimate how use of parks and playgrounds changes with air quality during wildfire season in the northwest United States. We find small reductions in park and playground visits on moderately polluted days, and large reductions, to 50-60% of baseline visits, when pollution levels are high. Disaggregating results by neighborhood characteristics, we find a significantly greater behavioral response to moderate levels of air pollution in neighborhoods with higher socio-economic status, although responses to high levels of pollution are similar across neighborhood types.
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The master tumor suppressor p53 activates transcription in response to various cellular stresses in part by facilitating recruitment of the transcription machinery to DNA. Recent studies have documented a direct yet poorly characterized interaction between p53 and RNA polymerase II (Pol II). Therefore, we dissected the human p53/Pol II interaction via single-particle cryo-electron microscopy, structural docking, and biochemical analyses. This study reveals that p53 binds Pol II via the Rpb1 and Rpb2 subunits, bridging the DNA-binding cleft of Pol II proximal to the upstream DNA entry site. In addition, the key DNA-binding surface of p53, frequently disrupted in various cancers, remains exposed within the assembly. Furthermore, the p53/Pol II cocomplex displays a closed conformation as defined by the position of the Pol II clamp domain. Notably, the interaction of p53 and Pol II leads to increased Pol II elongation activity. These findings indicate that p53 may structurally regulate DNA-binding functions of Pol II via the clamp domain, thereby providing insights into p53-regulated Pol II transcription.
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Modelos Moleculares , ARN Polimerasa II/química , ARN Polimerasa II/metabolismo , Proteína p53 Supresora de Tumor/química , Proteína p53 Supresora de Tumor/metabolismo , Microscopía por Crioelectrón , Exodesoxirribonucleasas/metabolismo , Humanos , Unión Proteica , Dominios Proteicos , Estructura Cuaternaria de Proteína , Elongación de la Transcripción GenéticaRESUMEN
In this study, we pilot a novel use of location quotient and proportional comparison methodology paired with mobile device location data. Specifically, we sought to understand visitation patterns in an urban park context based on visitor home locale socio-demographics, using an example from Fairmount Park in Philadelphia, PA, USA. We examined visitors' home locale demographics (i.e., percent African American and median household income) across a variety of park amenities (e.g., playgrounds, ball fields, splashpads), using location quotients and proportional analyses to compare the home locale demographics of visitors to specific amenities to park visitors as a whole. Findings illustrate amenities with over- or underrepresentation of visitors from certain socio-demographic groups, with bivariate analyses documenting intersectionality between race and income. Results of such analyses can aid park managers in understanding use of particular amenities and service gaps among historically underserved populations, and in turn, support more equitable resource allocation.
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Strategies to overcome resistance to FMS-like tyrosine kinase 3 (FLT3)-targeted therapy in acute myeloid leukemia (AML) are urgently needed. We identified autophagy as one of the resistance mechanisms, induced by hypoxia and the bone marrow microenvironment via activation of Bruton tyrosine kinase (BTK). Suppressing autophagy/BTK sensitized FLT3- mutated AML to FLT3 inhibitor-induced apoptosis. Furthermore, co-targeting FLT3/BTK/aurora kinases with a novel multikinase inhibitor CG-806 (luxeptinib) induced profound apoptosis in FLT3-mutated AML by co-suppressing FLT3/BTK, antagonizing autophagy, and causing leukemia cell death in FLT3-wildtype AML by aurora kinase-mediated G2/M arrest and polyploidy, in addition to FLT3 inhibition. Thus, CG-806 exerted profound anti-leukemia activity against AML regardless of FLT3 mutation status. CG-806 also significantly reduced AML burden and extended survival in an in vivo patient-derived xenograft leukemia murine model of FLT3 inhibitor-resistant FLT3-ITD/TKD double-mutant primary AML. Taken together, these findings indicate that CG-806 has a unique mechanistic action and pre-clinical activity, which is presently undergoing clinical evaluation in both FLT3 wildtype and mutant AML.
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Leucemia Mieloide Aguda , Tirosina Quinasa 3 Similar a fms , Humanos , Animales , Ratones , Agammaglobulinemia Tirosina Quinasa/genética , Tirosina Quinasa 3 Similar a fms/genética , Apoptosis , Línea Celular Tumoral , Puntos de Control de la Fase G2 del Ciclo Celular , Leucemia Mieloide Aguda/tratamiento farmacológico , Leucemia Mieloide Aguda/genética , Leucemia Mieloide Aguda/metabolismo , Mutación , Inhibidores de Proteínas Quinasas/farmacología , Inhibidores de Proteínas Quinasas/uso terapéutico , Autofagia , Microambiente TumoralRESUMEN
Cellular therapy development and manufacturing has focused on providing novel therapeutic cell-based products for various diseases. The International Organization for Standardization (ISO) has provided guidance on critical quality attributes (CQAs) that shall be considered when testing and releasing cellular therapeutic products. Cell count and viability measurements are two of the CQAs that are determined during development, manufacturing, testing, and product release. The ISO Cell Counting Standard Part 1 and 2 addressed the needs for improving the quality of cell counting results. However, there is currently no guidance on the qualification and selection of a fit-for-purpose cell viability detection method. In this work, we present strategies for the characterization and comparison of AO/PI and AO/DAPI staining methods using the heat-killed (HK) and low temperature/nutrient-deprived (LT/ND) cell death models to evaluate the comparability of cell viability measurements and identify potential causes of differences. We compared the AO/PI and AO/DAPI staining methods using HK and LT/ND-generated dead cells, investigated the staining time effects on cell viability measurements, and determined their viability linearity with different mixtures of live and dead cells. Furthermore, we validated AO/PI and AO/DAPI cell viability measurement with a long-term cell proliferation assay. Finally, we demonstrate a practical example of cell viability measurement comparison using AO/PI and AO/DAPI on antibiotic-selected transduced Jurkat and THP-1 cells to select a fit-for-purpose method for functional genomics screening. The proposed strategies may potentially enable scientists to properly characterize, compare, and select cell viability detection methods that are critical for cellular therapeutic product development and manufacturing.
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The Minnesota Multiphasic Personality Inventory-3 (MMPI-3) includes two self-concept-oriented scales: Self-Doubt (SFD), a measure of low self-esteem, and Self-Importance (SFI), a measure of beliefs that one has special attributes and abilities. Past research has demonstrated that SFD and SFI measure related but distinct constructs. The present study focused on explicating the meaning and clinical implications of low SFI scores. Using three clinical samples (private practice and community mental health and private practice neuropsychology clinics), we investigated whether the presence of interpretable low SFI scores (< 39 T) in the context of interpretable SFD elevations (≥ 65 T) is associated with distinctive MMPI-3 findings, and whether low SFI scores add clinically meaningful information in predicting relevant extra-test criteria. Consistent meaningful findings were obtained with respect to implications of low SFI scores for assessment of depression- and social engagement-related constructs. Additionally, the full range of SFI scores was meaningfully and negatively correlated with depressive disorder diagnoses and suicidal ideation but yielded very small correlations with suicide attempt and nonmeaningful correlations with diagnoses of Social Anxiety or Avoidant Personality Disorder. Hierarchical logistic regression analyses showed that SFI scores could meaningfully increment other related MMPI-3 scales in predicting diagnosed depressive disorders, albeit with small effect sizes.
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MMPI , Trastornos de la Personalidad , Humanos , Trastornos de la Personalidad/diagnóstico , Intento de Suicidio , Ideación Suicida , AutoimagenRESUMEN
This report provides an overview of the discussions, presentations, and consensus thinking from the Workshop on Smart Data Collection for CryoEM held at the New York Structural Biology Center on April 6-7, 2022. The goal of the workshop was to address next generation data collection strategies that integrate machine learning and real-time processing into the workflow to reduce or eliminate the need for operator intervention.
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Recolección de DatosRESUMEN
The recent introduction of slow vacuum filtration (SVF) technology has shown great promise for reproducibly creating high-quality, large-area aligned films of single-wall carbon nanotubes (SWCNTs) from solution-based dispersions. Despite clear advantages over other SWCNT alignment techniques, SVF remains in the developmental stages due to a lack of an agreed-upon alignment mechanism, a hurdle which hinders SVF optimization. In this work, the filter membrane surface is modified to show how the resulting SWCNT nematic order can be significantly enhanced. It is observed that directional mechanical grooving on filter membranes does not play a significant role in SWCNT alignment, despite the tendency for nanotubes to follow the groove direction. Chemical treatments to the filter membrane are shown to increase SWCNT alignment by nearly 1/3. These findings suggest that membrane surface structure acts to create a directional flow along the filter membrane surface that can produce global SWCNT alignment during SVF, rather serving as an alignment template.
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Nanotubos de Carbono , Nanotubos de Carbono/química , VacioRESUMEN
Tether proteins attach the endoplasmic reticulum (ER) to other cellular membranes, thereby creating contact sites that are proposed to form platforms for regulating lipid homeostasis and facilitating non-vesicular lipid exchange. Sterols are synthesized in the ER and transported by non-vesicular mechanisms to the plasma membrane (PM), where they represent almost half of all PM lipids and contribute critically to the barrier function of the PM. To determine whether contact sites are important for both sterol exchange between the ER and PM and intermembrane regulation of lipid metabolism, we generated Δ-super-tether (Δ-s-tether) yeast cells that lack six previously identified tethering proteins (yeast extended synatotagmin [E-Syt], vesicle-associated membrane protein [VAMP]-associated protein [VAP], and TMEM16-anoctamin homologues) as well as the presumptive tether Ice2. Despite the lack of ER-PM contacts in these cells, ER-PM sterol exchange is robust, indicating that the sterol transport machinery is either absent from or not uniquely located at contact sites. Unexpectedly, we found that the transport of exogenously supplied sterol to the ER occurs more slowly in Δ-s-tether cells than in wild-type (WT) cells. We pinpointed this defect to changes in sterol organization and transbilayer movement within the PM bilayer caused by phospholipid dysregulation, evinced by changes in the abundance and organization of PM lipids. Indeed, deletion of either OSH4, which encodes a sterol/phosphatidylinositol-4-phosphate (PI4P) exchange protein, or SAC1, which encodes a PI4P phosphatase, caused synthetic lethality in Δ-s-tether cells due to disruptions in redundant PI4P and phospholipid regulatory pathways. The growth defect of Δ-s-tether cells was rescued with an artificial "ER-PM staple," a tether assembled from unrelated non-yeast protein domains, indicating that endogenous tether proteins have nonspecific bridging functions. Finally, we discovered that sterols play a role in regulating ER-PM contact site formation. In sterol-depleted cells, levels of the yeast E-Syt tether Tcb3 were induced and ER-PM contact increased dramatically. These results support a model in which ER-PM contact sites provide a nexus for coordinating the complex interrelationship between sterols, sphingolipids, and phospholipids that maintain PM composition and integrity.
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Membrana Celular/metabolismo , Retículo Endoplásmico/metabolismo , Fosfolípidos/metabolismo , Esteroles/metabolismo , Lípidos/biosíntesis , Proteínas de la Membrana/metabolismo , Receptores de Esteroides/metabolismo , Proteínas de Saccharomyces cerevisiae/metabolismo , LevadurasRESUMEN
Outdoor recreation decision-making has received significant research interest over the last fifty years. In the context of campsite choice, this previous research has almost exclusively used stated preference data and aspatial methods to understand decision-making. This present research seeks to understand how recreationists reach decisions on the selection of campsites and what aspects of the recreational setting drive demand through an examination of a big dataset of revealed preference data using a spatial regression. Specifically, we examine which managerial, social, and ecological aspects of the setting influence demand for campsites in Zion National Park's (USA) Watchman Campground using reservation data from the Recreation Information Database (RIDB). Results indicate that price, access to electricity, ease of access, and proximity to the Virgin River are significantly predictive of demand. Study implications for park management, including campsite allocation and distributive justice, are provided. Additionally, implications for future research methodology, including the use of transaction-style big data in protected area management research, are discussed.
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Macrodatos , Recreación , Ríos , Análisis EspacialRESUMEN
Reliably determining the physical properties of ice (e.g., crystal structure, adhesion strength, interfacial state, and molecular orientation) has proven to be both challenging and highly dependent on experiment-specific conditions, including surface roughness, ice formation, water purity, and measurement method. Here, non-destructive measurements of single-layer graphene (SLG) interfaced with bulk ice are used to determine temperature-dependent, ice-induced strain and estimate ice-created strain elastic density in SLG. The use of SLG enables the precise study of interfacial strain by monitoring the 2D Raman mode. Upon ice formation, a clear, ≈2 cm-1 decrease in the 2D mode frequency is observed, which is ascribed to a 0.012% biaxial tensile shear strain at the ice-SLG interface. From this shear strain value, the ice-created SLG elastic strain energy density is estimated to be 2.4 µJ m-2 . In addition to these Raman strain measurements, intentionally ionized water is used to show that water-mediated charging of the SLG surface manifests itself in a distinctly different manner than ice-induced strain. Finally, the localized nature of the Raman probe is used to map SLG regions with and without ice, suggesting that this method cannot only determine ice-induced interfacial strain, but also correlate ice adhesion properties with surface roughness and topology.