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41.
S D Gupta R S Mehan T R Tansey H T Chen G Goping I Goldberg I Shechter 《Journal of lipid research》1999,40(9):1572-1584
Farnesyl diphosphate synthase (FPPS: EC2.5.1.10), a key enzyme in isoprenoid metabolic pathways, catalyzes the synthesis of farnesyl diphosphate (FPP) an intermediate in the biosynthesis of both sterol and non-sterol isoprenoid end products. The localization of FPPS to peroxisomes has been reported (Krisans, S. K., J. Ericsson, P. A. Edwards, and G. A. Keller. 1994. J. Biol. Chem. 269: 14165;-14169). Using indirect immunofluorescence and immunoelectron microscopic techniques we show here that FPPS is localized predominantly in the peroxisomes of rat hepatoma H35 cells. However, the partial release of 60;-70% of cellular FPPS activity is observed by selective permeabilization of these cells with digitonin. Under these conditions, lactate dehydrogenase, a cytosolic enzyme, is completely released whereas catalase, a known peroxisomal enzyme, is fully retained. Digitonin treatment of H35 cells differentially affects the release of other peroxisomal enzymes involved in isoprenoid metabolism. For instance, mevalonate kinase and phosphomevalonate kinase are almost totally released (95% and 91%, respectively), whereas 3-hydroxy-3-methylglutaryl-CoA reductase is fully retained. Indirect immunoflourescence studies indicate that FPPS is localized in peroxisomes of Chinese hamster ovary (CHO)-K1 cells but is dispersed in the cytosol of ZR-82 cells, a mutant that lacks peroxisomes. Unlike in H35 cells, FPPS is completely released upon digitonin permeabilization of CHO-K1 and ZR-82 cells. In contrast, under the same permeabilization conditions, catalase is fully retained in CHO-K1 cells but completely released from ZR-82 cells. These studies indicate that FPPS and other enzymes in the isoprenoid biosynthetic pathways, involved in the formation of FPP, are differentially associated with peroxisomes and may easily diffuse to the cytosol. Based on these observations, the significance and a possible regulatory model in the formation of isoprenoid end-products are discussed. 相似文献
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Sequence-specific adenylations and deadenylations accompany changes in the translation of maternal messenger RNA after fertilization of Spisula oocytes 总被引:19,自引:0,他引:19
A dramatic change in the pattern of protein synthesis occurs within ten minutes after fertilization of Spisula oocytes. This change is regulated entirely at the translational level. We have used DNA clones complementary to five translationally regulated messenger RNAs to follow shifts in mRNA utilization at fertilization and to characterize alterations in mRNA structure that accompany switches in translational activity in vivo. Four of the mRNAs studied are translationally inactive in the oocyte. After fertilization two of these mRNAs are completely recruited onto polysomes, and two are partially recruited. All four of these mRNAs have very short poly(A) tracts in the oocyte; after fertilization the poly(A) tails lengthen considerably. In contrast, a fifth mRNA, that encoding alpha-tubulin mRNA, is translated very efficiently in the oocyte and is rapidly lost from polysomes after fertilization. Essentially all alpha-tubulin mRNA in the oocyte is poly(A)+ and a large portion of this mRNA undergoes complete deadenylation after fertilization. These results reveal a striking relationship between changes in adenylation and translational activity in vivo. This correlation is not perfect, however. Evidence for and against a direct role for polyadenylation in regulating these translational changes is discussed. Changes in poly(A) tails are the only alterations in mRNA sizes that we have been able to detect. This indicates that, at least for the mRNAs studied here, translational activation is not due to extensive processing of larger translationally incompetent precursors. We have also isolated several complementary DNA clones to RNAs encoded by the mitochondrial genome. Surprisingly, the poly(A) tracts of at least two of the mitochondrial RNAs also lengthen in response to fertilization. 相似文献
44.
Isolation of thermophilic fungi from alligator nesting material 总被引:1,自引:0,他引:1
M R Tansey 《Mycologia》1973,65(3):594-601
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Susan Page Agata Hosciło Henk Wösten Jyrki Jauhiainen Marcel Silvius Jack Rieley Henk Ritzema Kevin Tansey Laura Graham Harri Vasander Suwido Limin 《Ecosystems》2009,12(6):888-905
Studies of restoration ecology are well established for northern peatlands, but at an early stage for tropical peatlands. Extensive peatland areas in Southeast Asia have been degraded through deforestation, drainage and fire, leading to on- and off-site environmental and socio-economic impacts of local to global significance. To address these problems, landscape-scale restoration measures are urgently required. This paper reviews and illustrates, using information from on-going trials in Kalimantan, Indonesia, the current state of knowledge pertaining to (i) land-cover dynamics of degraded peatlands, (ii) vegetation rehabilitation, (iii) restoration of hydrology, (iv) rehabilitation of carbon sequestration and storage, and (v) promotion of sustainable livelihoods for local communities. For a 4500 km2 study site in Central Kalimantan, Indonesia, we show a 78% reduction in forest cover between 1973 and 2003 and demonstrate that fire, exacerbated by drainage, is the principal driver of land-use change. Progressive vegetation succession follows infrequent, low-intensity fires, but repeated and high-intensity fires result in retrogressive succession towards non-forest communities. Re-wetting the peat is an important key to vegetation restoration and protection of remaining peat carbon stocks. The effectiveness of hydrological restoration is discussed and likely impacts on greenhouse gas emissions evaluated. Initial results indicate that raised water levels have limited short-term impact on reducing CO2 emissions, but could be critical in reducing fire risk. We conclude that successful restoration of degraded peatlands must be grounded in scientific knowledge, relevant to socio-economic circumstances, and should not proceed without the consent and co-operation of local communities. 相似文献
48.
Tansey EM 《Comptes rendus biologies》2006,329(5-6):419-425
In 1936 Sir Henry Dale of London and Professor Otto Loewi from Graz shared the Nobel Prize in Physiology or Medicine for their work on chemical neurotransmission. This paper uses much unpublished archival material to augment an examination of Dale's work, from his discovery of naturally occurring acetylcholine in 1913, through to evidence of its role as a neurotransmitter at autonomic ganglia, post-ganglionic parasympathetic nerve terminals and the neuromuscular junction. 相似文献
49.
Cheng Cui Bishwanath Chatterjee Thomas P. Lozito Zhen Zhang Richard J. Francis Hisato Yagi Lisa M. Swanhart Subramaniam Sanker Deanne Francis Qing Yu Jovenal T. San Agustin Chandrakala Puligilla Tania Chatterjee Terry Tansey Xiaoqin Liu Matthew W. Kelley Elias T. Spiliotis Adam V. Kwiatkowski Rocky Tuan Gregory J. Pazour Neil A. Hukriede Cecilia W. Lo 《PLoS biology》2013,11(11)
Planar cell polarity (PCP) regulates cell alignment required for collective cell movement during embryonic development. This requires PCP/PCP effector proteins, some of which also play essential roles in ciliogenesis, highlighting the long-standing question of the role of the cilium in PCP. Wdpcp, a PCP effector, was recently shown to regulate both ciliogenesis and collective cell movement, but the underlying mechanism is unknown. Here we show Wdpcp can regulate PCP by direct modulation of the actin cytoskeleton. These studies were made possible by recovery of a Wdpcp mutant mouse model. Wdpcp-deficient mice exhibit phenotypes reminiscent of Bardet–Biedl/Meckel–Gruber ciliopathy syndromes, including cardiac outflow tract and cochlea defects associated with PCP perturbation. We observed Wdpcp is localized to the transition zone, and in Wdpcp-deficient cells, Sept2, Nphp1, and Mks1 were lost from the transition zone, indicating Wdpcp is required for recruitment of proteins essential for ciliogenesis. Wdpcp is also found in the cytoplasm, where it is localized in the actin cytoskeleton and in focal adhesions. Wdpcp interacts with Sept2 and is colocalized with Sept2 in actin filaments, but in Wdpcp-deficient cells, Sept2 was lost from the actin cytoskeleton, suggesting Wdpcp is required for Sept2 recruitment to actin filaments. Significantly, organization of the actin filaments and focal contacts were markedly changed in Wdpcp-deficient cells. This was associated with decreased membrane ruffling, failure to establish cell polarity, and loss of directional cell migration. These results suggest the PCP defects in Wdpcp mutants are not caused by loss of cilia, but by direct disruption of the actin cytoskeleton. Consistent with this, Wdpcp mutant cochlea has normal kinocilia and yet exhibits PCP defects. Together, these findings provide the first evidence, to our knowledge, that a PCP component required for ciliogenesis can directly modulate the actin cytoskeleton to regulate cell polarity and directional cell migration. 相似文献
50.
Regulator of G protein signaling 10 (RGS10), a GTPase accelerating protein (GAP) for G alpha subunits, is a negative regulator of NF-κB in microglia. Here, we investigated the role of RGS10 in macrophages, a closely related myeloid-derived cell type. Features of classical versus alternative activation were assessed in Rgs10-/- peritoneal and bone marrow-derived macrophages upon LPS or IL-4 treatments, respectively. Our results showed that Rgs10-/- macrophages produced higher levels of pro-inflammatory cytokines including TNF, IL-1β and IL-12p70 in response to LPS treatment and exerted higher cytotoxicity on dopaminergic MN9D neuroblastoma cells. We also found that Rgs10-/- macrophages displayed a blunted M2 phenotype upon IL-4 priming. Specifically, Rgs10-/- macrophages displayed lower YM1 and Fizz1 mRNA levels as measured by QPCR compared to wild type macrophages upon IL-4 treatment and this response was not attributable to differences in IL-4 receptor expression. Importantly, phagocytic activities of Rgs10-/- macrophages were blunted in response to IL-4 priming and/or LPS treatments. However, there was no difference in chemotaxis between Rgs10-/- and WT macrophages. Our data indicate that Rgs10-/- macrophages displayed dysregulated M1 responses along with blunted M2 alternative activation responses, suggesting that RGS10 plays an important role in determining macrophage activation responses. 相似文献