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1.
Susan Aiston Matthew P Coghlan Loranne Agius 《European journal of biochemistry》2003,270(13):2773-2781
Multiple signalling pathways are involved in the mechanism by which insulin stimulates hepatic glycogen synthesis. In this study we used selective inhibitors of glycogen synthase kinase-3 (GSK-3) and an allosteric inhibitor of phosphorylase (CP-91149) that causes dephosphorylation of phosphorylase a, to determine the relative contributions of inactivation of GSK-3 and dephosphorylation of phosphorylase a as alternative pathways in the stimulation of glycogen synthesis by insulin in hepatocytes. GSK-3 inhibitors (SB-216763 and Li+) caused a greater activation of glycogen synthase than insulin (90% vs. 40%) but a smaller stimulation of glycogen synthesis (30% vs. 150%). The contribution of GSK-3 inactivation to insulin stimulation of glycogen synthesis was estimated to be less than 20%. Dephosphorylation of phosphorylase a with CP-91149 caused activation of glycogen synthase and translocation of the protein from a soluble to a particulate fraction and mimicked the stimulation of glycogen synthesis by insulin. The stimulation of glycogen synthesis by phosphorylase inactivation cannot be explained by either inhibition of glycogen degradation or activation of glycogen synthase alone and suggests an additional role for translocation of synthase. Titrations with the phosphorylase inactivator showed that stimulation of glycogen synthesis by insulin can be largely accounted for by inactivation of phosphorylase over a wide range of activities of phosphorylase a. We conclude that a signalling pathway involving dephosphorylation of phosphorylase a leading to both activation and translocation of glycogen synthase is a critical component of the mechanism by which insulin stimulates hepatic glycogen synthesis. Selective inactivation of phosphorylase can mimic insulin stimulation of hepatic glycogen synthesis. 相似文献
2.
Liu H Radhakrishnan P Magoun L Prabu M Campellone KG Savage P He F Schiffer CA Leong JM 《Molecular microbiology》2002,45(6):1557-1573
Attachment to host cells by enterohaemorrhagic Escherichia coli (EHEC) is associated with the formation of a highly organized cytoskeletal structure containing filamentous actin, termed an attaching and effacing (AE) lesion. Intimin, an outer membrane protein of EHEC, is required for the formation of AE lesions, as is Tir, a bacterial protein that is translocated into the host cell to function as a receptor for intimin. We established a yeast two-hybrid assay for intimin-Tir interaction and, after random mutagenesis, isolated 24 point mutants in intimin, which disrupted Tir recognition in this system. Analysis of 11 point mutants revealed a correlation between recognition of recombinant Tir and the ability to trigger AE lesions. Many of the mutations fell within a 50-residue region near the C-terminus of intimin. Alanine-scanning mutagenesis of this region revealed four residues (Ser890, Thr909, Asn916 and Asn927) that are critical for Tir recognition. Mapping the sequences of EHEC intimin and Tir onto the crystal structure of the intimin-Tir complex of enteropathogenic E. coli predicts that each of these four intimin residues lies at the intimin-Tir interface and contributes to a pocket that interacts with Ile298 of EHEC Tir. Thus, this genetic approach to intimin function both identified residues critical for Tir binding and demonstrated a correlation between the ability to bind Tir and the ability to trigger actin focusing. 相似文献
3.
Evamaria W. Koch M. Stephen Ailstock Dale M. Booth Deborah J. Shafer A. Dale Magoun 《Restoration Ecology》2010,18(4):584-595
We tested the hypothesis that currents, waves, and sediment grain size affect the dispersal of seeds and seedlings of the submersed angiosperms Ruppia maritima, Potamogeton perfoliatus and Stuckenia pectinata. Seed settling velocities and initiation of motion of seeds and seedlings and distance transported were quantified on four sediment types under a range of currents and waves in a flume. The rapid settling velocities of R. maritima and S. pectinata seeds and the increased settling velocity of P. perfoliatus in currents above 8 cm/second suggest that primary dispersal of these species is localized to the general area colonized by their parents. Once settled within a bed, seeds are exposed to weak currents and waves, and are likely to be subject to sediment deposition which may further limit dispersal. In contrast, in restoration projects, the absence of vegetation is likely to make seeds more vulnerable to grazing and transport, and may contribute to the lack of plant establishment. If seeds germinate without being buried, they are susceptible to secondary dispersal at relatively low current velocities and small wave heights due to the drag exerted on the cotyledon. Sand grains tend to stick to the seed coat and rootlet of P. perfoliatus seedlings, perhaps a mechanism to reduce the chances of being displaced following germination. These data reveal the close links between sediment, water flow, and submersed angiosperm seedling establishment; these parameters should be considered when using seeds for restoration of submersed angiosperms. 相似文献
4.
Green AR Aiston S Greenberg CC Freeman S Poucher SM Brady MJ Agius L 《The Journal of biological chemistry》2004,279(45):46474-46482
Expression of the glycogen-targeting protein PTG promotes glycogen synthase activation and glycogen storage in various cell types. In this study, we tested the contribution of phosphorylase inactivation to the glycogenic action of PTG in hepatocytes by using a selective inhibitor of phosphorylase (CP-91149) that causes dephosphorylation of phosphorylase a and sequential activation of glycogen synthase. Similar to CP-91194, graded expression of PTG caused a concentration-dependent inactivation of phosphorylase and activation of glycogen synthase. The latter was partially counter-acted by the expression of muscle phosphorylase and was not additive with the activation by CP-91149, indicating that it is in part secondary to the inactivation of phosphorylase. PTG expression caused greater stimulation of glycogen synthesis and translocation of glycogen synthase than CP-91149, and the translocation of synthase could not be explained by accumulation of glycogen, supporting an additional role for glycogen synthase translocation in the glycogenic action of PTG. The effects of PTG expression on glycogen synthase and glycogen synthesis were additive with the effects of glucokinase expression, confirming the complementary roles of depletion of phosphorylase a (a negative modulator) and elevated glucose 6-phosphate (a positive modulator) in potentiating the activation of glycogen synthase. PTG expression mimicked the inactivation of phosphorylase caused by high glucose and counteracted the activation caused by glucagon. The latter suggests a possible additional role for PTG on phosphorylase kinase inactivation. 相似文献
5.
6.
Hui Liu Loranne Magoun Steve Luperchio David B. Schauer & John M. Leong 《Molecular microbiology》1999,34(1):67-81
Enterohaemorrhagic Escherichia coli (EHEC) has emerged as an important agent of diarrhoeal disease. Attachment to host cells, an essential step during intestinal colonization by EHEC, is associated with the formation of a highly organized cytoskeletal structure containing filamentous actin, termed an attaching and effacing (A/E) lesion, directly beneath bound bacteria. The outer membrane protein intimin is required for the formation of this structure, as is Tir, a bacterial protein that is translocated into the host cell and is thought to function as a receptor for intimin. To understand intimin function better, we fused EHEC intimin to a homologous protein, Yersinia pseudotuberculosis invasin, or to maltose-binding protein. The N-terminal 539 amino acids of intimin were sufficient to promote outer membrane localization of the C-terminus of invasin and, conversely, the N-terminal 489 amino acids of invasin were sufficient to promote the localization of the C-terminus of intimin. The C-terminal 181 residues of intimin were sufficient to bind mammalian cells that had been preinfected with an enteropathogenic E. coli strain that expresses Tir but not intimin. Binding of intimin derivatives to preinfected cells correlated with binding to recombinant Tir protein. Finally, the 181-residue minimal Tir-binding region of intimin, when purified and immobilized on latex beads, was sufficient to trigger A/E lesions on preinfected mammalian cells. 相似文献
7.
Audrey J. Magoun Clinton D. Long Michael K. Schwartz Kristine L. Pilgrim Richard E. Lowell Patrick Valkenburg 《The Journal of wildlife management》2011,75(3):731-739
We developed an integrated system for photographing a wolverine's (Gulo gulo) ventral pattern while concurrently collecting hair for microsatellite DNA genotyping. Our objectives were to 1) test the system on a wild population of wolverines using an array of camera and hair-snag (C&H) stations in forested habitat where wolverines were known to occur, 2) validate our ability to determine identity (ID) and sex from photographs by comparing photographic data with that from DNA, and 3) encourage researchers and managers to test the system in different wolverine populations and habitats and improve the system design. Of the 18 individuals (10 M, 8 F) for which we obtained genotypes over the 2 years of our study, there was a 100% match between photographs and DNA for both ID and sex. The integrated system made it possible to reduce cost of DNA analysis by >74%. Integrating motion-detection cameras and hair snags provides a cost-effective technique for wildlife managers to monitor wolverine populations in remote habitats and obtain information on important population parameters such as density, survival, productivity, and effective population size. © 2011 The Wildlife Society. 相似文献
8.
Modelling broad‐scale wolverine occupancy in a remote boreal region using multi‐year aerial survey data
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Justina C. Ray Lucy G. Poley Audrey J. Magoun Cheryl‐Lesley B. Chetkiewicz F. Meg Southee F. Neil Dawson Chris Chenier 《Journal of Biogeography》2018,45(7):1478-1489
Aim
We used data from aerial surveys of wolverine tracks collected in seven winters over a 10‐year period (2003–2012) within a 574,287 km2 study area to evaluate the broad‐scale pattern of wolverine occurrence across a remote northern boreal forest region, identifying areas of high and low occupancy.Location
Northern Ontario, Canada.Taxon
Wolverine (Gulo gulo Linnaeus, 1758).Methods
We collected wolverine tracks and observations in 100‐km2 hexagonal survey units, making a total of 6,664 visits to 3,039 units, visiting each 1–9 times. We used hierarchical Bayesian occupancy modelling to model wolverine occurrence, and included covariates with the potential to affect detection and/or occupancy probability of wolverines.Results
we detected wolverines on 946 visits, 14.2% of total visits. Probability of detecting a wolverine varied among years and between the two ecozones in the study area. Wolverine occupancy was negatively related to two important covariates, the geographical coordinate Easting and thawing degree‐days. A site occupancy probability map indicated that wolverine occupancy probabilities were highest, and standard error lowest, in the western and northern portions of the study area.Main conclusions
The occupancy framework enabled us to use observation data from tracks of this elusive, wide‐ranging carnivore over a vast, remote area while explicitly considering detectability and spatial autocorrelation, yielding a map of probable wolverine distribution in northern Ontario that would not be possible using other methods of detection across a large region. With resource development pressures increasing in this globally significant region in the face of a changing climate, it is important to monitor changes in distribution of species like wolverines that have low population growth rates, large spatial requirements and sensitivity to human disturbance. This study demonstrates a relatively cost‐effective and non‐invasive alternative to monitoring based on wolverine harvest records, which have not been available since 2009 in Ontario due to changes in the provincial regulatory regime for this threatened species. 相似文献9.
Craig L. Gardner James P. Lawler Jay M. Ver Hoef Audrey J. Magoun Kalin A. Kellie 《The Journal of wildlife management》2010,74(8):1894-1903
ABSTRACT We determined wolverine (Gulo gulo) distribution and occurrence probabilities using aerial surveys and hierarchical spatial modeling in a 180,000-km2 portion of Interior Alaska, USA. During 8 February-12 March 2006, we surveyed 149 of 180 1,000-km2 sample units for wolverine tracks. We observed wolverine tracks in 99 (66.4%) sample units. Wolverine detection probability was ≥69% throughout the survey period. Posterior occurrence probabilities of whether a wolverine track occurred in a sample unit was dependent on survey timing, number of transects flown, number of neighboring sample units with detected tracks, percentage of the sample unit with elevation ≤305 m, and human influences. Our model indicated strong evidence of occurrence (>0.80) in 72% of the 180 survey units, strong evidence of absence (<0.20) in 12%, and weak evidence of occurrence or absence (0.20–0.80) in 16%. Wolverine area of occupancy made up 83% of the study area. Simulations illustrated that 2–4 survey routes were necessary for the survey technique to provide strong evidence of wolverine presence or absence in Interior Alaska if a track was not identified along the first route. The necessary number of survey routes depends on the occurrence probability in a sample unit. We provided managers with a map of wolverine distribution in Interior Alaska and an efficient and lower-cost method to detect coarse-scale changes in wolverine distribution. Our technique was effective in both Interior Alaska and Ontario, Canada, suggesting it would be effective throughout most of the boreal forest range of wolverines where tracks can be readily observed from the air. The technique requires a certain skill level in recognizing tracks; it is essential that tracks are identified correctly and training may be necessary depending on surveyor experience. 相似文献
10.
Susan M. Abdel-Rahman Takashi Sugita Gloria M. González David Ellis Michalis Arabatzis Loranne Vella-Zahra Calude Viguié-Vallanet Masataro Hiruma J. Steven Leeder Barry Preuett 《Mycopathologia》2010,169(1):1-13
Trichophyton tonsurans is a widely distributed pathogen that demonstrates a significant degree of genetic and phenetic heterogeneity. To date, the
degree of genetic relatedness among geographically segregated isolates has not been explored. This investigation evaluates
the extent of genetic variation among an international population of T. tonsurans isolates and examines the relatedness of isolates within and between countries. Molecular strain typing was performed on
198 isolates obtained from 14 countries. A mixed-marker strategy utilizing 27 sequence variations in 13 gene loci was applied
to all isolates and cluster analysis was performed to examine the relationship between strains. Phylogenetic analysis was
used to corroborate the findings of the cluster analysis with T. equinum strains serving as an out-group. In total, 47 distinct strain types were identified represented by seven clusters and one
singleton. There appeared to be a moderate degree of clustering among isolates obtained from North America, Asia and Australia,
although European isolates were uniformly distributed among the majority of clusters. The degree of genetic variation observed
in this study coupled with the geographic localization would support the argument for allopatric divergence within this species. 相似文献