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921.
A gene from the mesophilic bacterium Dehalococcoides ethenogenes encodes a novel mannosylglycerate synthase
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Empadinhas N Albuquerque L Costa J Zinder SH Santos MA Santos H da Costa MS 《Journal of bacteriology》2004,186(13):4075-4084
Mannosylglycerate (MG) is a common compatible solute found in thermophilic and hyperthermophilic prokaryotes. In this study we characterized a mesophilic and bifunctional mannosylglycerate synthase (MGSD) encoded in the genome of the bacterium Dehalococcoides ethenogenes. mgsD encodes two domains with extensive homology to mannosyl-3-phosphoglycerate synthase (MPGS, EC 2.4.1.217) and to mannosyl-3-phosphoglycerate phosphatase (MPGP, EC 3.1.3.70), which catalyze the consecutive synthesis and dephosphorylation of mannosyl-3-phosphoglycerate to yield MG in Pyrococcus horikoshii, Thermus thermophilus, and Rhodothermus marinus. The bifunctional MGSD was overproduced in Escherichia coli, and we confirmed the combined MPGS and MPGP activities of the recombinant enzyme. The optimum activity of the enzyme was at 50 degrees C. To examine the properties of each catalytic domain of MGSD, we expressed them separately in E. coli. The monofunctional MPGS was unstable, while the MPGP was stable and was characterized. Dehalococcoides ethenogenes cannot be grown sufficiently to identify intracellular compatible solutes, and E. coli harboring MGSD did not accumulate MG. However, Saccharomyces cerevisiae expressing mgsD accumulated MG, confirming that this gene product can synthesize this compatible solute and arguing for a role in osmotic adjustment in the natural host. We did not detect MGSD activity in cell extracts of S. cerevisiae. Here we describe the first gene and enzyme for the synthesis of MG from a mesophilic microorganism and discuss the possible evolution of this bifunctional MGSD by lateral gene transfer from thermophilic and hyperthermophilic organisms. 相似文献
922.
Borges N Marugg JD Empadinhas N da Costa MS Santos H 《The Journal of biological chemistry》2004,279(11):9892-9898
Rhodothermus marinus responds to fluctuations in the growth temperature and/or salinity by accumulating mannosylglycerate (MG). Two alternative pathways for the synthesis of MG have been identified in this bacterium: a single-step pathway and a two-step pathway. In this work, the genetic and biochemical characterization of the two-step pathway was carried out with the goal of understanding the function of the two pathways and their regulatory mechanisms. Mannosyl-3-phosphoglycerate synthase (MPGS) of the two-step pathway was purified from R. marinus. Sequence information led to the isolation of two contiguous genes, mpgs (encoding MPGS) and mpgp (encoding mannosyl-3-phosphoglycerate phosphatase). The recombinant MPGS had a low specific activity compared with other homologous MPGSs and contained approximately 30 additional residues at the C terminus. Truncation of this extension produced a protein with a 10-fold higher specific activity. Moreover, the activity of the complete MPGS was enhanced upon incubation with R. marinus cell extracts, and protease inhibitors abolished activation. Therefore, the C-terminal peptide of MPGS was identified as a regulatory site for short term control of MG synthesis in R. marinus. The control of gene expression by heat and osmotic stress was also studied; the level of mannosylglycerate synthase involved in the single-step pathway was selectively enhanced by heat stress, whereas MPGS was overproduced in response to osmotic stress. The concomitant changes in the level of MG were assessed as well. We conclude that the two alternative pathways for the synthesis of MG are differently regulated at the level of expression to play specific roles in the adaptation of R. marinus to two different types of aggression. This is the only example of pathway multiplicity being rationalized in terms of the need to respond efficiently to distinct environmental stresses. 相似文献
923.
924.
Eneqvist T Lundberg E Karlsson A Huang S Santos CR Power DM Sauer-Eriksson AE 《The Journal of biological chemistry》2004,279(25):26411-26416
Transthyretin (TTR) is an extracellular transport protein involved in the distribution of thyroid hormones and vitamin A. So far, TTR has only been found in vertebrates, of which piscine TTR displays the lowest sequence identity with human TTR (47%). Human and piscine TTR bind both thyroid hormones 3,5,3'-triiodo-l-thyronine (T(3)) and 3,5,3',5'-tetraiodo-l-thyronine (thyroxine, T(4)). Human TTR has higher affinity for T(4) than T(3), whereas the reverse holds for piscine TTR. X-ray structures of Sparus aurata (sea bream) TTR have been determined as the apo-protein at 1.75 A resolution and bound to ligands T(3) and T(4), both at 1.9 A resolution. The apo structure is similar to human TTR with structural changes only at beta-strand D. This strand forms an extended loop conformation similar to the one in chicken TTR. The piscine TTR.T(4) complex shows the T(4)-binding site to be similar but not identical to human TTR, whereas the TTR.T(3) complex shows the I3' halogen situated at the site normally occupied by the hydroxyl group of T(4). The significantly wider entrance of the hormone-binding channel in sea bream TTR, in combination with its narrower cavity, provides a structural explanation for the different binding affinities of human and piscine TTR to T(3) and T(4). 相似文献
925.
Gómez-Moutón C Lacalle RA Mira E Jiménez-Baranda S Barber DF Carrera AC Martínez-A C Mañes S 《The Journal of cell biology》2004,164(5):759-768
Spatially restricted activation of signaling molecules governs critical aspects of cell migration; the mechanism by which this is achieved nonetheless remains unknown. Using time-lapse confocal microscopy, we analyzed dynamic redistribution of lipid rafts in chemoattractant-stimulated leukocytes expressing glycosyl phosphatidylinositol-anchored green fluorescent protein (GFP-GPI). Chemoattractants induced persistent GFP-GPI redistribution to the leading edge raft (L raft) and uropod rafts of Jurkat, HL60, and dimethyl sulfoxide-differentiated HL60 cells in a pertussis toxin-sensitive, actin-dependent manner. A transmembrane, nonraft GFP protein was distributed homogeneously in moving cells. A GFP-CCR5 chimera, which partitions in L rafts, accumulated at the leading edge, and CCR5 redistribution coincided with recruitment and activation of phosphatidylinositol-3 kinase gamma in L rafts in polarized, moving cells. Membrane cholesterol depletion impeded raft redistribution and asymmetric recruitment of PI3K to the cell side facing the chemoattractant source. This is the first direct evidence that lipid rafts order spatial signaling in moving mammalian cells, by concentrating the gradient sensing machinery at the leading edge. 相似文献
926.
Dondi E Roué G Yuste VJ Susin SA Pellegrini S 《Journal of immunology (Baltimore, Md. : 1950)》2004,173(6):3740-3747
Type I IFNs (IFN-alphabeta) enhance immune responses, notably T cell-mediated responses, in part by promoting the functional activities of dendritic cells. In this study, we analyzed the direct impact of IFN-alpha on proliferative and apoptotic signals upon in vitro activation of human naive CD4+ T lymphocytes. We demonstrate that IFN-alpha protects T cells from the intrinsic mitochondrial-dependent apoptosis early upon TCR/CD28 activation. IFN-alpha acts by delaying entry of cells into the G1 phase of the cell cycle, as well as by increasing Bcl-2 and limiting Bax activation. Later, upon activation, T cells that were exposed to IFN-alpha showed increased levels of surface Fas associated with partially processed caspase-8, a key component of the extrinsic apoptotic pathway. Caspase-8 processing was augmented furthermore by Fas ligation. Overall, these findings support a model whereby IFN-alpha favors an enhanced clonal expansion, yet it sensitizes cells to the Ag-induced cell death occurring at the end of an immune response. These observations point to a complex role of type I IFN in regulating the magnitude of proliferation and survival of naive CD4+ T cells during primary response and underline how crucial could be the timing of exposure to this cytokine. 相似文献
927.
Ready JS Sampaio I Schneider H Vinson C Dos Santos T Turner GF 《Journal of evolutionary biology》2006,19(4):1139-1148
Laboratory mate choice experiments have confirmed species status for cichlid fish in the African Great Lakes that differ in colour and little else. Colour differences between allopatric populations of the South American cichlid genus Apistogramma are known for many species, yet the status of such populations has not been previously tested. Analysis of the genetic relationships and mate choice characteristics of populations previously described as Apistogramma caetei from eastern Amazonia indicates genetic differentiation into at least three allopatric lineages, which also show strong prezygotic isolation through female mate choice, confirming them as Biological species. If future studies confirm that this result is indicative of a general trend, the species richness of the South American cichlid fishes may presently be seriously underestimated. 相似文献
928.
Segmentation consists on the progressive formation of repetitive embryonic structures, named somites, which are formed from the most rostral part of the presomitic mesoderm. Somites are subdivided into anterior and posterior compartments and several genes are differentially expressed in either compartment. This has provided evidence for the importance of establishing the anterior-posterior polarity within each somite, which is critical for the correct segmented pattern of the adult vertebrate body. Although all somites appear morphologically similar, fate map studies have shown that the first 4 somites do not give rise to segmented structures, in contrast to more posterior ones. Moreover, in several somitogenesis-related mutants the anterior somites are not affected while posterior somites present clear defects or do not form at all. Altogether these data suggest relevant differences between rostral and caudal somites. In order to check for molecular differences between anterior and posterior somites, we have performed a detailed expression pattern analysis of several Notch signalling related genes. For the first time, we show that the somitic expression pattern profile is not the same along the anterior-posterior axis and that the differences are not observed always at the same somite level. 相似文献
929.
The structure of the prenylated flavanonol, floranol (1=(2R,3R)-3,5,7-trihydroxy-2-(2-hydroxyphenyl)-6-methoxy-8-(3-methylbut-2-enyl)-4H-1-benzopyran-4-one), isolated from the roots of Dioclea grandiflora (Fabaceae), was unambiguously determined by X-ray analysis. The compound was tested for vasorelaxant activity. In endothelium-containing aortic rings, floranol (1) induced a concentration-dependent vasodilator effect in vessels precontracted with 0.1 microM phenylephrine with an IC(50) value of 19.9+/-2.4 microM. The removal of endothelium or pretreatment of vessels with the NO-synthase inhibitor L-NAME did not change the IC(50) and E(max) values for floranol-induced vasorelaxation. We conclude that floranol (1) should be acting directly in the rat-aorta smooth muscle cells to produce its vasorelaxant effect. The structure-activity relationship was discussed in terms of the 3-D floranol structure determined by X-ray crystallography. 相似文献
930.
We describe a novel polymorphic Alu insertion (DXS225) on the human X chromosome (Xq21.3) embedded into an L1 retrotransposon. The DXS225 polymorphism was genotyped in 684 males from the CEPH Human Genome Diversity Panel. This insertion was found in all regions of the globe, suggesting that it took place before modern humans spread from Africa ca. 100,000 years ago. However, only one Amerindian population (Karitiana) showed this insertion allele, which may have been introduced by European admixture. Thus, it appears likely that the Alu insertion was absent from pre-Columbian America. Analysis of molecular variance worldwide demonstrated that 92.2% of the genetic variance was concentrated within populations. DXS225 is flanked by two microsatellites (DXS8114 and DXS1002), which are 86 kb apart and are in very strong linkage disequilibrium. The combination of a unique event polymorphism on the X chromosome in linkage disequilibrium with two rapidly evolving microsatellites should provide a useful tool for studies of human evolution. 相似文献