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101.
Male germ cell expression of the PAS domain kinase PASKIN and its novel target eukaryotic translation elongation factor eEF1A1. 总被引:1,自引:0,他引:1
Katrin Eckhardt Juliane Troger Jana Reissmann D?rthe M Katschinski Klaus F Wagner Petra Stengel Uwe Paasch Peter Hunziker Emanuela Borter Sandra Barth Philipp Schlafli Patrick Spielmann Daniel P Stiehl Gieri Camenisch Roland H Wenger 《Cellular physiology and biochemistry》2007,20(1-4):227-240
PASKIN links energy flux and protein synthesis in yeast, regulates glycogen synthesis in mammals, and has been implicated in glucose-stimulated insulin production in pancreatic beta-cells. Using newly generated monoclonal antibodies, PASKIN was localized in the nuclei of human testis germ cells and in the midpiece of human sperm tails. A speckle-like nuclear pattern was observed for endogenous PASKIN in HeLa cells in addition to its cytoplasmic localization. By yeast two-hybrid screening, we identified the multifunctional eukaryotic translation elongation factor eEF1A1 as a novel interaction partner of PASKIN. This interaction was mapped to the PAS A and kinase domains of PASKIN and to the C-terminus of eEF1A1 using mammalian two-hybrid and GST pull-down assays. Kinase assays, mass spectrometry and site-directed mutagenesis revealed PASKIN auto-phosphorylation as well as eEF1A1 target phosphorylation mainly but not exclusively at Thr432. Wild-type but not kinase-inactive PASKIN increased the in vitro translation of a reporter cRNA. Whereas eEF1A1 did not localize to the nucleus, it co-localizes with PASKIN to the cytoplasm of HeLa cells. The two proteins also showed a remarkably similar localization in the midpiece of the sperm tail. These data suggest regulation of eEF1A1 by PASKIN-dependent phosphorylation in somatic as well as in sperm cells. 相似文献
102.
103.
Sun B Eckhardt ER Shetty S van der Westhuyzen DR Webb NR 《Journal of lipid research》2006,47(8):1700-1713
Previous studies have suggested that HDL retroendocytosis may play a role in scavenger receptor class B type I (SR-BI)-dependent selective lipid uptake in a cell-specific manner. To investigate this possibility, we developed methods to quantitatively measure HDL uptake and resecretion in fibroblast (COS-7) and hepatocyte (HepG2) cells expressing exogenous SR-BI. Approximately 17% and 24% of HDL associated in an SR-BI-dependent manner with COS-7 and HepG2 cells, respectively, accumulates intracellularly after a 10 min incubation. To determine whether this intracellular HDL undergoes retroendocytosis, we developed a pulse-chase assay whereby internalized biotinylated (125)I-HDL(3) secreted from cells is quantitatively precipitated from cell supernatants using immobilized streptavidin. Our results show a rapid secretion of a portion of intracellular HDL from both cell types (representing 4-7% of the total cell-associated HDL) that is almost complete within 30 min (half-life approximately 10 min). In COS-7 cells, the calculated rate of HDL secretion ( approximately 0.5 ng HDL/mg/min) was >30-fold slower than the rate of SR-BI-dependent selective cholesteryl ester (CE) uptake ( approximately 17 ng HDL/mg/min), whereas the rate of release of HDL from the cell surface ( approximately 19 ng HDL/mg/min) was similar to the rate of selective CE uptake. Notably, the rate of SR-BI-dependent HDL resecretion in COS-7 and HepG2 cells was similar. BLT1, a compound that inhibits selective CE uptake, does not alter the amount of SR-BI-mediated HDL retroendocytosis in COS-7 cells. From these data, we conclude that HDL retroendocytosis in COS-7 and HepG2 cells is similar and that the vast majority of SR-BI-dependent selective uptake occurs at the cell surface in both cell types. 相似文献
104.
A substantial proportion of human genes contain tissue-specifically DNA-methylated regions (TDMRs). However, little is known about the evolutionary conservation of differentially methylated loci, how they evolve, and the signals that regulate them. We have studied TDMR conservation in the PLG and TBX gene families and in 32 pseudogene–parental gene pairs. Among the members of the recently evolved PLG gene family, 5′-UTR methylation is conserved and inversely correlated with the cognate gene expression, indicating as well a conserved regulatory role of DNA methylation. Conversely, many genes of the much older TBX family display complementary tissue-specific methylation, suggesting an epigenetic complementation in the evolution of this gene family. Similar to gene families, unprocessed pseudogenes arose from gene duplications and we found TDMR conservation in some pseudogene–parental gene pairs displaying short evolutionary distances. However, for the majority of unprocessed pseudogenes and for all processed pseudogenes examined, we found that tissue-specific methylation arose de novo after gene duplication. 相似文献
105.
Cara L. Eckhardt Linda S. Adair Benjamin Caballero Josephine Avila Igor Y. Kon Jinzhong Wang Barry M. Popkin 《Obesity (Silver Spring, Md.)》2003,11(12):1553-1562
Objective: The goal was to assess the ability of BMI to predict body fat (BF) among youths in four countries and identify the degree to which additional anthropometric measures improve this prediction. BMI is widely recommended as an indicator of overweight. However, whether BMI adequately estimates BF and has the same meaning in different ethnic groups and youths has been questioned. Research Methods and Procedures: Data come from 456 Filipino, Chinese, Russian, and black South African youths, 6 to 16 years old. Percentage BF and fat mass index (FMI) were estimated by the deuterium dilution method. Skinfold thicknesses (triceps, subscapular, and suprailiac) and weight and height measures were collected. Percentage BF was regressed first on BMI and age and then with the addition of the skinfold measures. Linear models were run separately by country and sex. The models were repeated with FMI as the outcome. Results: The R2 values from the percentage BF models ranged from 0.13 to 0.69 in the first models to 0.38 to 0.81 in the full models. The values were lowest among Russian males ≥ 13 years and Russian females ≥ 13 years of age in the reduced and full models, respectively, and were highest among Chinese females. Using FMI as the outcome did not meaningfully change the results. Discussion: The ability of BMI to adequately predict BF and the additional predictivity of anthropometric measures varied widely across the samples, making its uniform use as a proxy for BF in youths from different countries questionable. 相似文献
106.
Because photosynthetic rates in C4 plants are the same at normal levels of O2 (c, 20 kPa) and at c, 2 kPa O2 (a conventional test for evaluating photorespiration in C3 plants) it has been thought that C4 photosynthesis is O2 insensitive. However, we have found a dual effect of O2 on the net rate of CO2 assimilation among species representing all three C4 subtypes from both monocots and dicots. The optimum O2 partial pressure for C4 photosynthesis at 30 °C, atmospheric CO2 level, and half full sunlight (1000 μmol quanta m?2 s?1) was about 5–10 kPa. Photosynthesis was inhibited by O2 below or above the optimum partial pressure. Decreasing CO2 levels from ambient levels (32.6 Pa) to 9.3 Pa caused a substantial increase in the degree of inhibition of photosynthesis by supra-optimum levels of O2 and a large decrease in the ratio of quantum yield of CO2 fixation/quantum yield of photosystem II (PSII) measured by chlorophyll a fluorescence. Photosystem II activity, measured from chlorophyll a fluorescence analysis, was not inhibited at levels of O2 that were above the optimum for CO2 assimilation, which is consistent with a compensating, alternative electron How as net CO2 assimilation is inhibited. At suboptimum levels of O2, however, the inhibition of photosynthesis was paralleled by an inhibition of PSII quantum yield, increased state of reduction of quinone A, and decreased efficiency of open PSII centres. These results with different C4 types suggest that inhibition of net CO2 assimilation with increasing O2 partial pressure above the optimum is associated with photorespiration, and that inhibition below the optimum O2 may be caused by a reduced supply of ATP to the C4 cycle as a result of inhibition of its production photochemically. 相似文献
107.
108.
大鼠蓝斑内注入谷氨酸钠的心血管效应及其中枢机制 总被引:5,自引:0,他引:5
本工作在乌拉坦麻醉、箭毒化、人工呼吸的大鼠观察到:(1)将 L-谷氨酸钠(Glu)微量注入蓝斑(LC)引起血压升高,心率无明显变化;注入 LC 邻近区引起血压降低、心率减慢。(2)在下丘脑的室旁核尾侧断脑可衰减 LC 加压效应,而室旁核头侧断脑对 LC 加压反应无明显影响,双侧延髓头端腹外侧区(RVL)内分别注射酚妥拉明、心得安、阿托品均使兴奋 LC 引起的加压效应衰减;提示蓝斑加压效应由室旁核和 RVL(及其内的α-、β-肾上腺素能受体,M-胆碱能受体)介导。 相似文献
109.
A rapid, simple, and inexpensive method has been developed for preparing UDP-N-acetylgalactosamine in amounts sufficient for several thousand assays of enzymes that employ this nucleotide sugar as substrate. The UDP-N-acetylglucosamine-4-epimerase in extracts of porcine submaxillary glands was used to convert UDP-N-acetylglucosamine to an equilibrium mixture of UDP-N-acetylglucosamine and UDP-N-acetylgalactosamine (molar ratio, 77:23). The two nucleotide sugars were separated from components in the extract by ion-exchange chromatography and then separated from one another by affinity chromatography on a column of Griffonia simplicifolia lectin I bound to agarose. The UDP-N-acetylgalactosamine was obtained in pure form by ion-exchange chromatography in an overall yield of 91% from the equilibrium mixture. The separation of the two nucleotide sugars by affinity chromatography also provides a rapid assay for the UDPGlcNAc-4-epimerase, which is more accurate and less time consuming than earlier published assays. 相似文献
110.
The H1 histones from erythrocytes, livers, intestines, testes, and embryos of Xenopus laevis have been examined electrophoretically. This species has been found to contain at least five electrophoretically resolvable lysine-rich histones in addition to the presumptive H5 histone of erythrocytes. Quantitative and qualitative distinctions between the H1 histones from each source were readily observed. Three H1 histones (H1A, H1B, and H1C) were found in both embryos and adult tissues, although in varying amounts. Two other H1 histones (H1D and H1E) were found only in adult tissues. Comparative SDS gel V8 protease cleavage maps of the lysine-rich histones from testes and erythrocytes have demonstrated that the “adult-specific” H1D and H1E are not artifacts of proteolysis and may be closely related to the presumptive H5 histone. Spermatogenic cells were found to be similar to embryonic cells in being deficient in H1D and H1E. These observations suggest that H1D and H1E are enriched in cell types with low rates of cell division similar to the mammalian H1° histone. The results presented here demonstrate a previously unrecognized degree of developmental and cell-specific variance in the H1 histones of Xenopus laevis. 相似文献