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1.
2.
Lichenicolous fungi are a species-rich biological group growing on lichen thalli. Here, we analyze the genetic structure of the lichenicolous basidiomycete Tremella lobariacearum and three host species (Lobaria pulmonaria, Lobaria macaronesica, and Lobaria immixta) in Macaronesia. We used ordination and analysis of molecular variance to investigate the structuring of genetic variation, and a simulation test to investigate whether rDNA haplotypes of T. lobariacearum were significantly associated with host species. To investigate the evolutionary and demographic history of the lichenicolous fungus and its hosts, we used coalescent samplers to generate trees, and Bayesian skyline plots. We found that the hosts were most important in structuring populations of the lichenicolous species. Despite their wide geographic distribution, the same haplotypes of T. lobariacearum consistently associated with a given host species. Our results suggest that the Lobaria hosts create a selective environment for the lichenicolous fungus. Both the pathogen and the host populations exhibited substantial genetic structure. However, evolutionary and demographic histories differed between the parasite and its hosts, as evidenced by different divergence times and tree topologies. 相似文献
3.
Molybdenum is an essential component of the cofactors of many metalloenzymes including nitrate reductase and Mo-nitrogenase. The cyanobacterium Anabaena variabilis ATCC 29413 uses nitrate and atmospheric N2 as sources of nitrogen for growth. Two of the three nitrogenases in this strain are Mo-dependent enzymes, as is nitrate reductase; thus, transport of molybdate is important for growth of this strain. High-affinity transport of molybdate in A. variabilis was mediated by an ABC-type transport system encoded by the products of modA and modBC. The modBC gene comprised a fused orf including components corresponding to modB and modC of Escherichia coli. The deduced ModC part of the fused gene lacked a recognizable molybdate-binding domain. Expression of modA and modBC was induced by starvation for molybdate. Mutants in modA or modBC were unable to grow using nitrate or Mo-nitrogenase. Growth using the alternative V-nitrogenase was not impaired in the mutants. A high concentration of molybdate (10 microM) supported normal growth of the modBC mutant using the Nif1 Mo-nitrogenase, indicating that there was a low-affinity molybdate transport system in this strain. The modBC mutant did not detectably transport low concentrations of 99Mo (molybdate), but did transport high concentrations. However, such transport was observed only after cells were starved for sulphate, suggesting that an inducible sulphate transport system might also serve as a low-affinity molybdate transport system in this strain. 相似文献
4.
Phosphatidylinositol-3,5-Bisphosphate is a potent and selective inhibitor of acid sphingomyelinase 总被引:4,自引:0,他引:4
Kölzer M Arenz C Ferlinz K Werth N Schulze H Klingenstein R Sandhoff K 《Biological chemistry》2003,384(9):1293-1298
Acid sphingomyelinase (A-SMase, EC 3.1.4.12) catalyzes the lysosomal degradation of sphingomyelin to phosphorylcholine and ceramide. Inherited deficiencies of acid sphingomyelinase activity result in various clinical forms of Niemann-Pick disease, which are characterised by massive lysosomal accumulation of sphingomyelin. Sphingomyelin hydrolysis by both, acid sphingomyelinase and membrane-associated neutral sphingomyelinase, plays also an important role in cellular signaling systems regulating proliferation, apoptosis and differentiation. Here, we present a potent and selective novel inhibitor of A-SMase, L-alpha-phosphatidyl-D-myo-inositol-3,5-bisphosphate (PtdIns3,5P2), a naturally occurring substance detected in mammalian, plant and yeast cells. The inhibition constant Ki for the new A-SMase inhibitor PtdIns3,5P2 is 0.53 microM as determined in a micellar assay system with radiolabeled sphingomyelin as substrate and recombinant human A-SMase purified from insect cells. Even at concentrations of up to 50 microM, PtdIns3,5P2 neither decreased plasma membrane-associated, magnesium-dependent neutral sphingomyelinase activity, nor was it an inhibitor of the lysosomal hydrolases beta-hexosaminidase A and acid ceramidase. Other phosphoinositides tested had no or a much weaker effect on acid sphingomyelinase. Different inositol-bisphosphates were studied to elucidate structure-activity relationships for A-SMase inhibition. Our investigations provide an insight into the structural features required for selective, efficient inhibition of acid sphingomyelinase and may also be used as starting point for the development of new potent A-SMase inhibitors optimised for diverse applications. 相似文献
5.
Goats and some sheep synthesize a juvenile hemoglobin, Hb C (alpha 2 beta
C2), at birth and produce this hemoglobin exclusively during severe anemia.
Sheep that synthesize this juvenile hemoglobin are of the A haplotype.
Other sheep, belonging to a separate group, the B haplotype, do not
synthesize hemoglobin C and during anemia continue to produce their adult
hemoglobin. To understand the basis for this difference we have determined
the structural organization of the beta- globin locus of B-type sheep by
constructing and isolating overlapping genomic clones. These clones have
allowed us to establish the linkage map 5' epsilon I-epsilon II-psi beta
I-beta B-epsilon III-epsilon IV- psi beta II-beta F3' in this haplotype.
Thus, B sheep lack four genes, including the BC gene, and have only eight
genes, compared with the 12 found in the goat globin locus. The goat
beta-globin locus is as follows: 5' epsilon I-epsilon II-psi beta X-beta
C-epsilon III-epsilon IV-psi beta Z-beta A-epsilon V-epsilon VI-psi beta
Y-beta F3'. Southern blot analysis of A-type sheep reveals that these
animals have a beta- globin locus similar to that of goat, i.e., 12 globin
genes. Thus, the beta-globin locus of B-haplotype sheep resembles that of
cows and may have retained the duplicated locus of the ancestor of cows and
sheep. Alternatively, the B-sheep locus arrangement may be the result of a
deletion of a four-gene set from the triplicated locus.
相似文献
6.
7.
Lakshmiprasad Potluri Aneta Karczmarek Jolanda Verheul Andre Piette Jean‐Marc Wilkin Nadine Werth Manuel Banzhaf Waldemar Vollmer Kevin D. Young Martine Nguyen‐Distèche Tanneke Den Blaauwen 《Molecular microbiology》2010,77(2):300-323
The distribution of PBP5, the major D,D‐carboxypeptidase in Escherichia coli, was mapped by immunolabelling and by visualization of GFP fusion proteins in wild‐type cells and in mutants lacking one or more D,D‐carboxypeptidases. In addition to being scattered around the lateral envelope, PBP5 was also concentrated at nascent division sites prior to visible constriction. Inhibiting PBP2 activity (which eliminates wall elongation) shifted PBP5 to midcell, whereas inhibiting PBP3 (which aborts divisome invagination) led to the creation of PBP5 rings at positions of preseptal wall formation, implying that PBP5 localizes to areas of ongoing peptidoglycan synthesis. A PBP5(S44G) active site mutant was more evenly dispersed, indicating that localization required enzyme activity and the availability of pentapeptide substrates. Both the membrane bound and soluble forms of PBP5 converted pentapeptides to tetrapeptides in vitro and in vivo, and the enzymes accepted the same range of substrates, including sacculi, Lipid II, muropeptides and artificial substrates. However, only the membrane‐bound form localized to the developing septum and restored wild‐type rod morphology to shape defective mutants, suggesting that the two events are related. The results indicate that PBP5 localization to sites of ongoing peptidoglycan synthesis is substrate dependent and requires membrane attachment. 相似文献
8.
We present a multiplex real-time PCR assay for the simultaneous identification of three morphologically similar species of lichen-forming fungi, Lobaria pulmonaria, Lobaria immixta and Lobaria macaronesica. Based on TaqMan MGB (minor groove binding) probes targeting the fungal internal transcribed spacer (ITS nrDNA) region, our assay unambiguously identifies known samples from all the three species, thus providing a powerful alternative to the more expensive DNA-sequencing techniques. 相似文献
9.
Limited autolysis reduces the Ca2+ requirement of a smooth muscle Ca2+-activated protease 总被引:5,自引:0,他引:5
Chicken gizzard smooth muscle contains large amounts of Ca2+-activated protease activity. Approximately 15 mg of purified enzyme can be obtained from 1 kg of fresh muscle. The enzyme consists of two subunits (Mr = 80,000 and 30,000) present in a 1:1 molar ratio. In the presence of CaCl2, the 80,000/30,000-dalton heterodimer (form I) is rapidly converted by limited autolysis to a 76,000/18,000-dalton species (form II). Both the 80,000- and 30,000-dalton subunits are degraded simultaneously. Moreover, the Ca2+ dependence for autolysis (K0.5 = 300 microM) is identical for both subunits. Neither the time course nor the Ca2+ dependence of the autolytic conversion reaction is altered by 10- and 20-fold molar excesses of substrate. Limited autolysis markedly reduces the Ca2+ requirement for substrate degradation. Using N-[ethyl-2-3H]maleimide-labeled 27,000-dalton cardiac myosin light chains as substrate, the Ca2+ requirement of form I was found to be quite high (K0.5 = 150 microM). Under similar conditions, the Ca2+ requirement of form II was 30-fold lower (K0.5 = 5 microM). Limited autolysis did not alter the specific activity of the enzyme. Our results demonstrate that smooth muscle contains an abundant amount of Ca2+-activated protease. Moreover, autolysis of this enzyme may play an important regulatory role by converting the native form to a species that is fully active at physiological levels of intracellular calcium ion. 相似文献
10.
Ultraviolet (UV) light alters cutaneous structure and function. Prior work has shown loss of dermal hyaluronan after UV-irradiation of human skin, yet UV exposure increases total glycosaminoglycan (GAG) content in mouse models. To more fully describe UV-induced alterations to cutaneous GAG content, we subjected human volunteers to intermediate-term (5 doses/week for 4 weeks) or single-dose UV exposure. Total dermal uronyl-containing GAGs increased substantially with each of these regimens. We found that UV exposure substantially increased dermal content of chondroitin sulfate (CS), but not hyaluronan, heparan sulfate, or dermatan sulfate. UV induced the accumulation of both the 4-sulfated (C4S) and 6-sulfated (C6S) isoforms of CS, but in distinct distributions. Next, we examined several CS proteoglycan core proteins and found a significant accumulation of dermal and endothelial serglycin, but not of decorin or versican, after UV exposure. To examine regulation in vitro, we found that UVB in combination with IL-1α, a cytokine upregulated by UV radiation, induced serglycin mRNA in cultured dermal fibroblasts, but did not induce the chondroitin sulfate synthases. Overall, our data indicate that intermediate-term and single-dose UVB exposure induces specific GAGs and proteoglycan core proteins in human skin in vivo. These molecules have important biologic functions and contribute to the cutaneous response to UV. 相似文献