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Shivani Patel Donna McKeon Kimheak Sao Changsong Yang Nicole M. Naranjo Tatyana M. Svitkina Ryan J. Petrie 《Molecular biology of the cell》2021,32(7):579
Human fibroblasts can switch between lamellipodia-dependent and -independent migration mechanisms on two-dimensional surfaces and in three-dimensional (3D) matrices. RhoA GTPase activity governs the switch from low-pressure lamellipodia to high-pressure lobopodia in response to the physical structure of the 3D matrix. Inhibiting actomyosin contractility in these cells reduces intracellular pressure and reverts lobopodia to lamellipodial protrusions via an unknown mechanism. To test the hypothesis that high pressure physically prevents lamellipodia formation, we manipulated pressure by activating RhoA or changing the osmolarity of the extracellular environment and imaged cell protrusions. We find RhoA activity inhibits Rac1-mediated lamellipodia formation through two distinct pathways. First, RhoA boosts intracellular pressure by increasing actomyosin contractility and water influx but acts upstream of Rac1 to inhibit lamellipodia formation. Increasing osmotic pressure revealed a second RhoA pathway, which acts through nonmuscle myosin II (NMII) to disrupt lamellipodia downstream from Rac1 and elevate pressure. Interestingly, Arp2/3 inhibition triggered a NMII-dependent increase in intracellular pressure, along with lamellipodia disruption. Together, these results suggest that actomyosin contractility and water influx are coordinated to increase intracellular pressure, and RhoA signaling can inhibit lamellipodia formation via two distinct pathways in high-pressure cells. 相似文献
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A series of 10 synthetic peptides containing varying degrees of charge and hydrophobicity was used to study the effects of peptide composition and buffer pH on the selectivity of separations by capillary zone electrophoresis (CZE). A simple model is used to explain the effect of buffer pH on the separation. It was found that pH is an important parameter affecting the selectivity of CZE separations. Furthermore, it is shown that the selectivity of the separation is such that peptides differing in neutral amino acid composition can be resolved, and that even differences in a peptide's amino acid sequence can be detected. A protease digest of beta-lactoglobulin A is shown as a practical example of a separation of a complex peptide mixture. 相似文献
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Mariêve Picard Ryan J. Petrie Judith Antoine-Bertrand Etienne Saint-Cyr-Proulx Jose-France Villemure Nathalie Lamarche-Vane 《Cellular signalling》2009,21(12):1961-1973
Netrin-1 attracts or repels growing axons during development. The UNC5 receptors mediate the repulsive response, either alone or in complex with DCC receptors. The signaling mechanisms activated by UNC5 are poorly understood. Here, we examined the role of Rho GTPases in UNC5a signaling. We found that UNC5a induced neurite outgrowth in N1E-115 neuroblastoma cells in a netrin-1- and Rac1-dependent manner. UNC5a lacking its cytoplasmic tail also mediated this effect. In fibroblasts, UNC5a was able to activate RhoA and to a lower extent Rac1 and Cdc42 in response to netrin-1. Using Fluorescence Resonance Energy Transfer (FRET) intermolecular probes, we visualized the spatial and temporal activation of Rac1, Cdc42 and RhoA in live N1E-115 cells expressing UNC5a during neurite outgrowth. We found that Rac1 but not Cdc42 was transiently activated at the leading edge of the cell during neurite initiation. However, at later times when well-developed neurites were formed, active RhoA was found in the cell body and at the base of the neuronal leading process in UNC5a-expressing cells. Together, these findings demonstrate that the netrin-1 receptor UNC5a is able to induce neurite outgrowth and to differentially activate RhoA and Rac1 during neurite extension in a spatial and temporal manner. 相似文献
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Matthew D. Petrie Debra P. C. Peters N. Dylan Burruss Wenjie Ji Heather M. Savoy 《Global Change Biology》2019,25(10):3305-3318
Dryland vegetation is influenced by biotic and abiotic land surface template (LST) conditions and precipitation (PPT), such that enhanced vegetation responses to periods of high PPT may be shaped by multiple factors. High PPT stochasticity in the Chihuahuan Desert suggests that enhanced responses across broad geographic areas are improbable. Yet, multiyear wet periods may homogenize PPT patterns, interact with favorable LST conditions, and in this way produce enhanced responses. In contrast, periods containing multiple extreme high PPT pulse events could overwhelm LST influences, suggesting a divergence in how climate change could influence vegetation by altering PPT periods. Using a suite of stacked remote sensing and LST datasets from the 1980s to the present, we evaluated PPT‐LST‐Vegetation relationships across this region and tested the hypothesis that enhanced vegetation responses would be initiated by high PPT, but that LST favorability would underlie response magnitude, producing geographic differences between wet periods. We focused on two multiyear wet periods; one of above average, regionally distributed PPT (1990–1993) and a second with locally distributed PPT that contained two extreme wet pulses (2006–2008). 1990–1993 had regional vegetation responses that were correlated with soil properties. 2006–2008 had higher vegetation responses over a smaller area that were correlated primarily with PPT and secondarily to soil properties. Within the overlapping PPT area of both periods, enhanced vegetation responses occurred in similar locations. Thus, LST favorability underlied the geographic pattern of vegetation responses, whereas PPT initiated the response and controlled response area and maximum response magnitude. Multiyear periods provide foresight on the differing impacts that directional changes in mean climate and changes in extreme PPT pulses could have in drylands. Our study shows that future vegetation responses during wet periods will be tied to LST favorability, yet will be shaped by the pattern and magnitude of multiyear PPT events. 相似文献
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Griffin MR Monto AS Belongia EA Treanor JJ Chen Q Chen J Talbot HK Ohmit SE Coleman LA Lofthus G Petrie JG Meece JK Hall CB Williams JV Gargiullo P Berman L Shay DK;U.S. Flu-VE Network 《PloS one》2011,6(8):e23085
We estimated the effectiveness of four monovalent pandemic influenza A (H1N1) vaccines (three unadjuvanted inactivated, one live attenuated) available in the U.S. during the pandemic. Patients with acute respiratory illness presenting to inpatient and outpatient facilities affiliated with four collaborating institutions were prospectively recruited, consented, and tested for influenza. Analyses were restricted to October 2009 through April 2010, when pandemic vaccine was available. Patients testing positive for pandemic influenza by real-time RT-PCR were cases; those testing negative were controls. Vaccine effectiveness was estimated in logistic regression models adjusted for study community, patient age, timing of illness, insurance status, enrollment site, and presence of high-risk medical conditions. Pandemic virus was detected in 1,011 (15%) of 6,757 enrolled patients. Fifteen (1%) of 1,011 influenza positive cases and 1,042 (18%) of 5,746 test-negative controls had record-verified pandemic vaccination >14 days prior to illness onset. Adjusted effectiveness (95% confidence interval) for pandemic vaccines combined was 56% (23%, 75%). Adjusted effectiveness for inactivated vaccines alone (79% of total) was 62% (25%, 81%) overall and 32% (-92%, 76%), 89% (15%, 99%), and -6% (-231%, 66%) in those aged 0.5 to 9, 10 to 49, and 50+ years, respectively. Effectiveness for the live attenuated vaccine in those aged 2 to 49 years was only demonstrated if vaccination >7 rather than >14 days prior to illness onset was considered (61%∶ 12%, 82%). Inactivated non-adjuvanted pandemic vaccines offered significant protection against confirmed pandemic influenza-associated medical care visits in young adults. 相似文献