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  • 1
    Publication Date: 2022-05-26
    Description: © The Author(s), 2019. This article is distributed under the terms of the Creative Commons Attribution License. The definitive version was published in Kim, S., Kalappurakkal, J. M., Mayor, S., & Rosen, M. K. Phosphorylation of nephrin induces phase separated domains that move through actomyosin contraction. Molecular Biology of the Cell, 30(24), (2019): 2996–3012, doi:10.1091/mbc.E18-12-0823.
    Description: The plasma membrane of eukaryotic cells is organized into lipid and protein microdomains, whose assembly mechanisms and functions are incompletely understood. We demonstrate that proteins in the nephrin/Nck/N-WASP actin-regulatory pathway cluster into micron-scale domains at the basal plasma membrane upon triggered phosphorylation of transmembrane protein nephrin. The domains are persistent but readily exchange components with their surroundings, and their formation is dependent on the number of Nck SH3 domains, suggesting they are phase separated polymers assembled through multivalent interactions among the three proteins. The domains form independent of the actin cytoskeleton, but acto-myosin contractility induces their rapid lateral movement. Nephrin phosphorylation induces larger clusters at the cell periphery, which are associated with extensive actin assembly and dense filopodia. Our studies illustrate how multivalent interactions between proteins at the plasma membrane can produce micron-scale organization of signaling molecules, and how the resulting clusters can both respond to and control the actin cytoskeleton.
    Description: We thank Hongtao Yu (University of Texas Southwestern Medical Center [UTSW]) for providing the HeLa cell line used in this work; Dan Billadeau and Timothy Gomez (Mayo Clinic) for providing antibodies; Nico Stuurman (University of California, San Francisco) for assistance with STORM imaging; Kate Luby-Phelps and Abhijit Bugde (UTSW Live Cell Imaging Core Facility) for their assistance in epifluorescence and spinning disk confocal experiments; Sudeep Banjade for advice on designing the S3, S2, S1 constructs; Khuloud Jaqaman (UTSW) for advice on cluster motility analysis; Salman Banani and Jonathan Ditlev (UTSW) for critical reading of the manuscript; and members of the Rosen lab and participants in the MBL/HHMI Summer Institutes for advice and helpful discussions. This work was supported by a Howard Hughes Medical Institute Collaborative Innovation Award; the Welch Foundation (I-1544 to M.K.R.); a J.C. Bose Fellowship from the Department of Science and Technology, government of India (to S.M.); a Margadarshi Fellowship from the Wellcome Trust—Department of Biotechnology, India Alliance (IA/M/15/1/502018 to S.M.). Research in the Rosen lab is supported by the Howard Hughes Medical Institute.
    Repository Name: Woods Hole Open Access Server
    Type: Article
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  • 2
    Electronic Resource
    Electronic Resource
    Oxford, UK : Blackwell Publishing Ltd
    Journal of management studies 25 (1988), S. 0 
    ISSN: 1467-6486
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Economics
    Type of Medium: Electronic Resource
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  • 3
    Electronic Resource
    Electronic Resource
    Oxford, UK : Blackwell Publishing Ltd
    Annals of the New York Academy of Sciences 533 (1988), S. 0 
    ISSN: 1749-6632
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Natural Sciences in General
    Type of Medium: Electronic Resource
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  • 4
    Electronic Resource
    Electronic Resource
    s.l. : American Chemical Society
    Environmental science & technology 22 (1988), S. 398-405 
    ISSN: 1520-5851
    Source: ACS Legacy Archives
    Topics: Chemistry and Pharmacology , Energy, Environment Protection, Nuclear Power Engineering
    Type of Medium: Electronic Resource
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  • 5
    ISSN: 1520-6882
    Source: ACS Legacy Archives
    Topics: Chemistry and Pharmacology
    Type of Medium: Electronic Resource
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  • 6
    Electronic Resource
    Electronic Resource
    Oxford, UK : Blackwell Publishing Ltd
    Sedimentology 35 (1988), S. 0 
    ISSN: 1365-3091
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Geosciences
    Notes: Gravity cores of Holocene sediments from a shallow ephemeral lake in the Coorong region (Pellet Lake, southeastern coastal Australia) show a mineral assemblage and sequence particular to its hydrology. The mineralogical sequence above an initial dolomitic siliciclastic sand reflects conditions of increasing salinity in the lower portions of the core (i.e. organic-rich aragonite to magnesite + hydromagnesite + aragonite) followed by a relative decrease in salinity (i.e. magnesite + aragonite + hydromagnesite to aragonite + hydromagnesite) in the upper portions of the core. This sequence is capped by ˜ 0.4 m of micritic dolomite and minor amounts of hydromagnesite, with the relative abundance of dolomite increasing upwards. Three stratigraphically and spatially distinct dolomite units (upper, lower and margin) are recognized using stable carbon and oxygen isotope data, unit cell calculations and MgCO3 mole per cent data of the dolomite.Detailed X-ray diffraction (XRD) analyses of samples with more than 80% dolomite shows that the dolomite is ordered. Average unit cell parameters, calculated from the XRD patterns, indicate that the upper dolomite unit has crystal lattices expanded in the co direction (co= 16.09 Å) relative to ideal dolomite (co= 16.02 Å) and contracted in the ao direction (ao= 4.796 Å) relative to ideal dolomite (ao= 4.812 Å). The mol fraction of MgCO3 in the upper dolomite shows up to 4.0 ±M 2.0 mole per cent excess Mg in the dolomite crystal lattice (calculated from XRD). This unusual dolomite crystal chemistry is probably generated by rapid precipitation from solutions which have greatly elevated Mg/Ca ratios. Transmission electron microscopy reveals that the upper dolomite has a heterogeneous microstructure which also suggests rapid precipitation from solution. The modulated microstructure found in calcium-rich dolomite is completely lacking. Dolomite ordering reflections are present in electron diffraction patterns, but are weak.Stable oxygen and carbon isotope values of the upper dolomite are tightly grouped (ave. δ18O ∼+ 7.55%o, δ13C ∼+ 4.10%o), yet show three upward-lightening oxygen cycles. The oxygen cycles correlate with three upward decreases in the calculated Mg content of the dolomite zone. These cycles may indicate the increased importance of rain-water dilution of the brine at times when the water in the lake was at its shallowest levels.Analyses of the lower dolomite and the margin dolomite suggest that these units precipitated more slowly from less evaporitic brines than the upper dolomite unit. The lower dolomite is close to stoichiometric, has less evaporitic stable isotope values than the upper dolomite, and has only a slightly expanded co-axis. The margin dolomite is Ca-rich, has a more homogeneous microstructure, and has expanded ao and co axes.The abundance of relatively soluble Mg-bearing phases, such as hydromagnesite and magnesite, may supply additional magnesium for the dolomitization of aragonite and calcite during subsequent diagenesis and burial of the sediment. This process may leave a finely laminated dolomicrite deposit which retains little, if any, evidence of evaporite minerals.
    Type of Medium: Electronic Resource
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  • 7
    ISSN: 1745-6584
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Energy, Environment Protection, Nuclear Power Engineering , Geosciences
    Notes: The combined use of geophysical (time domain reflectometry [TDR]), isotopic (δ18 O and δ2H) and chemical (NO 3 -N, Cl) techniques indicates that residence times in the unsaturated zone of a fractured basalt aquifer in the Pukekohe region of the North Island, New Zealand, are at least six months. Each technique provides useful information on specific aspects of recharge and residence times, but combined they provide a basis for determining the mechanisms of water movement in the unsaturated zone. TDR soil moisture measurements provide similar rainfall recharge estimates as model calculations over longer time periods (monthly or seasonal) but may be more accurate for shorter time scales (weekly or daily).The TDR measurements indicate that recharge occurs mainly in the late autumn and winter months. Measurements of nitrate concentrations in the soil profile over this same period, suggest that nitrate is mineralized in the soil during the summer and is flushed past the root zone at the beginning of the recharge period, in early winter. Nitrate concentrations in the soil profile do not increase in concentration later in the recharge period, even after significant recharge events have occurred, because all stored nitrate has already been released. This pattern of nitrate movement indicates that fertilizer applications in spring and early summer will leach less to ground water and applications made in autumn and winter will leach more. Estimations of yearly recharge from mean monthly chloride concentrations (730 mm/yr) are roughly in agreement with the TDR estimates (680 mm/year). However, there is a large range in the chloride estimates (〉100%) because monthly chloride concentration measurements in both the rain and soil water vary due to the proximity of the site to the ocean.
    Type of Medium: Electronic Resource
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  • 8
    ISSN: 1476-4687
    Source: Nature Archives 1869 - 2009
    Topics: Biology , Chemistry and Pharmacology , Medicine , Natural Sciences in General , Physics
    Notes: [Auszug] The Rho-family GTP-hydrolysing proteins (GTPases), Cdc42, Rac and Rho, act as molecular switches in signalling pathways that regulate cytoskeletal architecture, gene expression and progression of the cell cycle. Cdc42 and Rac transmit many signals through GTP-dependent binding to effector proteins ...
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  • 9
    ISSN: 1476-4687
    Source: Nature Archives 1869 - 2009
    Topics: Biology , Chemistry and Pharmacology , Medicine , Natural Sciences in General , Physics
    Notes: [Auszug] The Rho GDP-dissociation inhibitors (GDIs) negatively regulate Rho-family GTPases,. The inhibitory activity of GDI derives both from an ability to bind the carboxy-terminal isoprene of Rho family members and extract them from membranes,, and from inhibition of GTPase cycling between the GTP- ...
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  • 10
    ISSN: 1476-4687
    Source: Nature Archives 1869 - 2009
    Topics: Biology , Chemistry and Pharmacology , Medicine , Natural Sciences in General , Physics
    Notes: [Auszug] To investigate the possible interaction of pTyr221 with the Crk SH2 domain, full-length mCrk, and an amino-terminal fragment truncated immediately after the first SH3 domain (mCrk23, resi-dues 1-197) were expressed in bacteria. Figure 1 illustrates a gradient, enhanced-sensitivity 'H-15N ...
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