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31.
32.
Yeast Saccharomyces cerevisiae MTO2, MTO1, and MSS1 genes encoded highly conserved tRNA modifying enzymes for the biosynthesis of carboxymethylaminomethyl (cmnm)5s2U34 in mitochondrial tRNALys, tRNAGlu, and tRNAGln. In fact, Mto1p and Mss1p are involved in the biosynthesis of the cmnm5 group (cmnm5U34), while Mto2p is responsible for the 2-thiouridylation (s2U34) of these tRNAs. Previous studies showed that partial modifications at U34 in mitochondrial tRNA enabled mto1, mto2, and mss1 strains to respire. In this report, we investigated the functional interaction between MTO2, MTO1, and MSS1 genes by using the mto2, mto1, and mss1 single, double, and triple mutants. Strikingly, the deletion of MTO2 was synthetically lethal with a mutation of MSS1 or deletion of MTO1 on medium containing glycerol but not on medium containing glucose. Interestingly, there were no detectable levels of nine tRNAs including tRNALys, tRNAGlu, and tRNAGln in mto2/mss1, mto2/mto1, and mto2/mto1/mss1 strains. Furthermore, mto2/mss1, mto2/mto1, and mto2/mto1/mss1 mutants exhibited extremely low levels of COX1 and CYTB mRNA and 15S and 21S rRNA as well as the complete loss of mitochondrial protein synthesis. The synthetic enhancement combinations likely resulted from the completely abolished modification at U34 of tRNALys, tRNAGlu, and tRNAGln, caused by the combination of eliminating the 2-thiouridylation by the mto2 mutation with the absence of the cmnm5U34 by the mto1 or mss1 mutation. The complete loss of modifications at U34 of tRNAs altered mitochondrial RNA metabolisms, causing a degradation of mitochondrial tRNA, mRNA, and rRNAs. As a result, failures in mitochondrial RNA metabolisms were responsible for the complete loss of mitochondrial translation. Consequently, defects in mitochondrial protein synthesis caused the instability of their mitochondrial genomes, thus producing the respiratory-deficient phenotypes. Therefore, our findings demonstrated a critical role of modifications at U34 of tRNALys, tRNAGlu, and tRNAGln in maintenance of mitochondrial genome, mitochondrial RNA stability, translation, and respiratory function. 相似文献
33.
A weak chemiluminescence (CL) emission was observed due to the production of singlet oxygen ((1)O(2)) during the decomposition of peroxomonosulphate (HSO(5)(-)) catalysed by cobalt(II). Low molecular mass aliphatic monocarboxylic acids, such as formic, acetic, propionic, butyric and valeric acids, influenced the CL emission, and the reaction of aliphatic monocarboxylic acids with HSO(5)(-)/Co(2+) solution was further investigated using a flow injection analysis (FIA) CL method. The results indicated that the CL intensities of aliphatic monocarboxylic acids were improved with increase in the carbon chain length in the potassium peroxomonosulphate-cobalt(II) sulphate system. Generation of singlet oxygen was confirmed by the fact that the CL emission of aliphatic monocarboxylic acids with the HSO(5)(-)/Co(2+) solution was quenched by NaN(3), and from the CL spectrum of the reaction system. Additionally, a possible mechanism of aliphatic monocarboxylic acids CL emission enhancement was proposed. 相似文献
34.
Haishan Tian Lu Tang Yi Wang Xiaojie Wang Lili Guan Jian Zhang Xiaoping Wu Xiaokun Li 《International journal of peptide research and therapeutics》2011,17(2):123-129
Recombinant human keratinocyte growth factor-2 (rhKGF-2) has previously been expressed in Escherichia coli using isopropyl-β-d-thiogalactopyranoside (IPTG), a non-metabolizable and expensive compound, as the inducer. In order to determine whether IPTG
could be replaced with the cheap and natural lactose to induce rhKGF-2 expression, we examined the expression of rhKGF-2 in
flask culture and 30-l fermentation using lactose as the inducer. The optimized fermentation induced with lactose resulted
in 1,382 g of cell mass, corresponding to a 84% enhancement in cell mass compared with IPTG induction. While the expression
level of rhKGF-2 induced with lactose was comparable to that induced with IPTG, the solubility of target protein was increased
by lactose induction than by IPTG induction. The recombinant protein was further purified by cation exchange and heparin-affinity
chromatography. 255 milligrams of pure rhKGF-2 was achieved per liter culture by lactose induction, 52% higher than that obtained
by IPTG induction. A preliminary biochemical characterization of purified rhKGF-2 was performed by Western blotting and mitogenic
activity analysis, and the results demonstrated that the purified lactose-induced rhKGF-2 could react with anti-human KGF-2
antibody and stimulate the proliferation of FGFR2-IIIb-transfected mouse BaF3 cells as IPTG-induced rhKGF-2 could do. 相似文献
35.
Cowan JW Wang X Guan R He K Jiang J Baumann G Black RA Wolfe MS Frank SJ 《The Journal of biological chemistry》2005,280(19):19331-19342
Growth hormone receptor (GHR) is a cytokine receptor superfamily member that binds growth hormone (GH) via its extracellular domain and signals via interaction of its cytoplasmic domain with JAK2 and other signaling molecules. GHR is a target for inducible metalloprotease-mediated cleavage in its perimembranous extracellular domain, a process that liberates the extracellular domain as the soluble GH-binding protein and leaves behind a cell-associated GHR remnant protein containing the transmembrane and cytoplasmic domains. GHR metalloproteolysis can be catalyzed by tumor necrosis factor-alpha-converting enzyme (ADAM-17) and is associated with down-modulation of GH signaling. We now study the fate of the GHR remnant protein. By anti-GHR cytoplasmic domain immunoblotting, we observed that the remnant induced in response to phorbol ester or platelet-derived growth factor has a reliable pattern of appearance and disappearance in both mouse preadipocytes endogenously expressing GHR and transfected fibroblasts expressing rabbit GHR. Lactacystin, a specific proteasome inhibitor, did not appreciably change the time course of remnant appearance or clearance but allowed detection of the GHR stub, a receptor fragment slightly smaller than the remnant but containing the C terminus of the remnant (receptor cytoplasmic domain). In contrast, MG132, another (less specific) proteasome inhibitor, strongly inhibited remnant clearance and prevented stub appearance. Inhibitors of gamma-secretase, an aspartyl protease, also prevented the appearance of the stub, even in the presence of lactacystin, and concomitantly inhibited remnant clearance in the same fashion as MG132. In addition, mouse embryonic fibroblasts derived from presenilin 1 and 2 (PS1/2) knockouts recapitulated the gamma-secretase inhibitor studies, as compared with their littermate controls (PS1/2 wild type). Confocal microscopy indicated that the GHR cytoplasmic domain became localized to the nucleus in a fashion dependent on PS1/2 activity. These data indicate that the GHR is subject to sequential proteolysis by metalloprotease and gamma-secretase activities and may suggest GH-independent roles for the GHR. 相似文献
36.
Naus S Reipschläger S Wildeboer D Lichtenthaler SF Mitterreiter S Guan Z Moss ML Bartsch JW 《Biological chemistry》2006,387(3):337-346
ADAM proteases are type I transmembrane proteins with extracellular metalloprotease domains. As for most ADAM family members, ADAM8 (CD156a, MS2) is involved in ectodomain shedding of membrane proteins and is linked to inflammation and neurodegeneration. To identify potential substrates released under these pathologic conditions, we screened 10-mer peptides representing amino acid sequences from extracellular domains of various membrane proteins using the ProteaseSpot system. A soluble ADAM8 protease containing a pro- and metalloprotease domain was expressed in E. coli and purified as active protease owing to autocatalytic prodomain removal. From 34 peptides tested in the peptide cleavage assay, significant cleavage by soluble ADAM8 was observed for 14 peptides representing membrane proteins with functions in inflammation and neurodegeneration, among them the beta-amyloid precursor protein (APP). The in vivo relevance of the ProteaseSpot method was confirmed by cleavage of full-length APP with ADAM8 in human embryonic kidney 293 cells expressing tagged APP. ADAM8 cleaved APP with similar efficiency as ADAM10, whereas the inactive ADAM8 mutant did not. Exchanging amino acids at defined positions in the cleavage sequence of myelin basic protein (MBP) revealed sequence criteria for ADAM8 cleavage. Taken together, the results allowed us to identify novel candidate substrates that could be cleaved by ADAM8 in vivo under pathologic conditions. 相似文献
37.
Biocomplexity theory is becoming increasingly important in understanding natural vegetation dynamics and interrelation among all components of the ecosystem. In this study, based on the field investigation of plant species and environmental factors (altitude, microtopography, soil water content, and soil nutrients) in an arid valley of the upper reaches of Minjiang River, Sichuan Province, southwestern China, plant community complexity and its relationship with environmental factors, community diversity, species evenness and richness were studied. Both total and structural complexities of the communities showed a “high- low-high” tendency with the increase in altitude of the area, which meant that the complexity of communities was the highest at the sites of low and high altitude, whereas it was the lowest at the sites of intermediate altitude. It was found that the total community complexity had significant quadratic correlations with soil organic matter (SOM) content, total nitrogen (N), hydrolyzable N, soil water content, and available potassium (K), whereas it had no significant correlations with soil total K, total phosphorus (P), available P, and pH value. The total community complexity positively correlated with community diversity, species evenness and species richness, whereas the structural complexity negatively correlated with the community evenness. Of the two components of the total community complexity, namely, the structural complexity and the structural diversity, the structural complexity was more sensitive than the structural diversity to the changes of species in the community, which was not only related to the community evenness but also to the community richness. The relative contribution of both the structural complexity and the structural diversity to the total complexity would be different for different study areas or ecosystems. 相似文献
38.
Litterfall Production Along Successional and Altitudinal Gradients of Subtropical Monsoon Evergreen Broadleaved Forests in Guangdong, China 总被引:3,自引:0,他引:3
Guoyi Zhou Lili Guan Xiaohua Wei Deqiang Zhang Qianmei Zhang Junhua Yan Dazhi Wen Juxiu Liu Shuguang Liu Zhongliang Huang Guohui Kong Jiangming Mo Qingfa Yu 《Plant Ecology》2007,188(1):77-89
Evaluation of litterfall production is important for understanding nutrient cycling, forest growth, successional pathways, and interactions with environmental variables in forest ecosystems. Litterfall was intensively studied during the period of 1982–2001 in two subtropical monsoon vegetation gradients in the Dinghushan Biosphere Reserve, Guangdong Province, China. The two gradients include: (1) a successional gradient composed of pine forest (PF), mixed pine and broadleaved forest (MF) and monsoon evergreen broadleaved forest (BF), and (2) an altitudinal gradient composed of Baiyunci ravine rain forest (BRF), Qingyunci ravine rain forest (QRF), BF and mountainous evergreen broadleaved forest (MMF). Mean annual litterfall production was 356, 861 and 849 g m−2 for PF, MF and BF of the successional gradient, and 1016, 1061, 849 and 489 g m−2 for BRF, QRF, BF and MMF of the altitudinal gradient, respectively. As expected, mean annual litterfall of the pioneer forest PF was the lowest, but rapidly increased over the observation period while those in other forests were relatively stable, confirming that forest litterfall production is closely related to successional stages and growth patterns. Leaf proportions of total litterfall in PF, MF, BF, BRF, QRF and MMF were 76.4%, 68.4%, 56.8%, 55.7%, 57.6% and 69.2%, respectively, which were consistent with the results from studies in other evergreen broadleaved forests. Our analysis on litterfall monthly distributions indicated that litterfall production was much higher during the period of April to September compared to other months for all studied forest types. Although there were significant impacts of some climate variables (maximum and effective temperatures) on litterfall production in some of the studied forests, the mechanisms of how climate factors (temperature and rainfall) interactively affect litterfall await further study. 相似文献
39.
花椒属植物生物碱研究进展 总被引:3,自引:0,他引:3
花椒属植物全世界约有250种,我国约有39种14变种。近年来研究发现花椒属植物生物碱具有多种药理作用与临床功能,现就近20年来花椒属生物碱和花椒生物碱的提取、分离、测定方法以及其药理作用等研究作一综述。 相似文献
40.