Metallothioneins have been isolated from Saccharomyces cerevisiae CUP1 mutants generated by Wright et al. (Wright, C. F., Hamer, D. H., and McKenney, K. (1986) Nucleic Acids Res. 14, 8489-8499). In the mutant metallothioneins, pairs of cysteinyl residues have been converted to seryl residues. The mutant proteins differ only in the positions of the double substitutions; each mutant molecule contains 10 cysteinyl residues. Each mutant protein lacks the first 8 residues at the amino terminus from the decoded gene sequence of the CUP1 locus. Mutant molecules consist of 53 residues analogous to the wild-type metallothionein and are designated 9/11, 24/26, 36/38, and 49/50 (in reference to the sequence positions of the Cys----Ser conversions). The properties of the mutant metallothioneins are vastly different, and host cells harboring the different plasmid-encoded mutant molecules show marked differences in sensitivity to CuSO4. Growth inhibition was observed at CuSO4 concentrations up to mM in cells containing the 9/11, 24/26, and 36/38 molecules, but not for cells containing protein 49/50. A CuSO4 concentration of 5 mM was required to inhibit the growth of yeast containing either 49/50 or the wild-type metallothionein. In the purified proteins the copper binding stoichiometry of each molecule, except protein 24/26, was nearly 8 mol eq. Protein 24/26 bound 5.5 copper ions/molecule. The Cu(I) chelator bathocuproine disulfonate reacted with over 50% of the copper ions in proteins 9/11, 24/26, and 36/38, but less than 10% of the copper ions in proteins 49/50 and wild-type metallothionein were reactive. The thiolates in 9/11, 24/26, and 36/38 were also more reactive in a disulfide exchange reaction with dithiodipyridine compared with the sulfhydryls in 49/50 and the wild-type molecules. The four mutant copper proteins are luminescent and exhibit a similar quantum yield. The cluster structures contributing to the particular electronic transitions are markedly more sensitive to oxygen in proteins 9/11, 24/26, and 36/38 compared with 49/50 and the wild-type molecules. The air-sensitive proteins exhibit a tertiary fold not recognized by polyclonal antibodies directed to a conformational epitope on yeast Cu-metallothionein. Protein 49/50 cross-reacts with the antibody in a concentration-dependent fashion similar to the wild-type protein. Mutation of 2 cysteinyl residues in the carboxyl portion of metallothionein does not significantly alter properties of the molecule, whereas mutation of several cysteines in the amino-terminal portion of the molecule yields a different conformation. 相似文献
The HLA haplotype segregation and autoantibody spectrum in 7 type I (insulin-dependent) diabetic multiplex families of North Indian origin were determined. Of the total of 17 diabetic sibs, 7 shared both haplotypes and 3 shared one haplotype with the proband. No HLA-non-identical sibs were observed. This distribution of haplotypes was non-random (P approximately equal to 0.005). The mode of inheritance was compatible with an autosomal recessive model, while a dominant model was unlikely. Pancreatic islet-cell antibodies were found in 23.5% of affected sibs, but in no healthy family member. A high incidence of other autoantibodies (parietal-cell and thyroglobulin/thyroid microsomal antibodies) was detected in both the diabetic patients (26.3%), and in healthy first-degree relatives (22.2%). These findings emphasize the role of HLA-linked genes and autoimmunity in the pathogenesis of type I diabetes in North India. 相似文献
The immune response in leprosy forms a spectrum with lepromatous leprosy patients exhibiting specific unresponsiveness to antigens of Mycobacterium leprae. This unresponsiveness is thought to be related to the prevalence of T8-positive lymphocyte in these lepromatous lesions. To analyze the immunoregulatory function of these T8 cells, we developed simple procedures to extract lymphocytes from skin biopsy specimens of patients with leprosy. These lymphocytes were sorted for T8 and T4 positive cells, and cell lines were established by expansion with interleukin 2 (IL 2) and irradiated feeder cells. All T8 positive lines tested were positive for IL 2 receptors and HLA-DR determinants. These lines were additionally assayed for lepromin-induced suppression of the normal peripheral blood lymphocyte Con A proliferative response. Thirteen of 32 lines from six lepromatous patients showed significant suppressor activity, whereas nine lines from six tuberculoid patients and one line from normal peripheral blood failed to show suppression (p less than 0.001). Taken together, the finding of M. leprae-triggered suppressor cells within lepromatous skin lesions may in part explain the M. leprae unresponsiveness of lepromatous leprosy patients. 相似文献
Summary
Ruellia tweediana and R. tuberosa are large flowered chasmogamous diploids (n=17) with normal meiosis and fertility. F1 hybrids, successful in only one direction (R. tweediana x R. tuberosa), are vegetatively vigorous and possess 17 often heteromorphic bivalents with high degree of segregational irregularities. It is exclusively cleistogamous and completely pollen and seed sterile. Like F1, the artificial amphidiploid (n=34) is also cleistogamous but shows preferential chromosome pairing with complete restoration of fertility. The parental chromosomes are sufficiently differentiated and cleistogamy is either genie or due to gene-cytoplasm interaction but sterility is entirely chromosomal. All floral parts excepting calyx are highly deformed. Such a deformity is associated with sterility in the F1 but with fertility in the amphidiploid. This is perhaps the first case of origin by hybridization of a true breeding and fully fertile cleistogamous taxon from two chasmogamous species. It also shows the extent and nature of change in breeding system brought about by hybridization and/or polyploidy.The chromosome numbers in the six, out of 16, obligate cleistogamous taxa (Table 4) show that they are high polyploids. Perhaps their origin has been in the same manner as in the present case.
Zusammenfassung
Ruellia tweediana und R. tuberosa sind großblütige, chasmogame Diploide (n=17) mit normaler Meiosis und Fertilität. Die F1-Hybriden, die nur in einer Richtung gelingen (R. tweediana x R. tuberosa), sind vegetativ kräftig und besitzen häufig 17 heteromorphe Bivalente mit einem hohen Anteil an Spaltungsunregelmäßigkeiten. Die Hybride ist ausschließlich kleistogam und vollkommen pollen- und samensteril. Wie die F1 ist auch die künstlich hergestellte Amphidiploide (n=34) kleistogam und zeigt eine präferentielle Chromosomenpaarung mit völliger Wiederherstellung der Fertilität. Die elterlichen Chromosomen sind genügend differenziert. Die Kleistogamie ist entweder genisch bedingt oder auf eine Gen-Cytoplasma-Interaktion zurückzuführen, die Sterilität ist ausschließlich durch die Chromosomen verursacht. Alle Teile der Blüte mit Ausnahme der Calyx sind stark deformiert. Bei der F1 ist diese Deformation mit Sterilität verbunden, die amphidiploide Form ist jedoch fertil. Das ist vielleicht der erste Fall eines aus der Kreuzung zweier chasmogamer Spezies hervorgegangenen reinerbigen und voll fertilen kleistogamen Taxons. Es läßt sich auch der Umfang und die Art der durch Hybridisierung und durch Polyploidie verursachten Änderung des Zuchtsystems erkennen. Die Chromosomenzahl bei 6 von 16 obligaten kleistogamen Taxa (Tab. 4) zeigt, daß sie hochpolyploid sind. Vielleicht sind sie auf eine gleiche Weise wie im vorliegenden Falle entstanden.
Purine hydroxylase II from Aspergillus nidulans has been purified to near homogeneity. The enzyme has a pI of 5.7, a molecular weight of 300,000, and two subunits with molecular weight of 153,000 each. The enzyme contains 2 FAD, 2 molybdenum atoms, and 4 (2 Fe-2S) iron-sulfur centers per molecule and exhibits broad specificity for reducing and oxidizing substrates. Among the more notable characteristics are the ability to oxidize hypoxanthine and nicotinic acid but not xanthine and virtually complete inactivity with oxygen. Moreover, while the enzyme is inactivated by borate and methanol, it is very resistant to cyanide and arsenite and it not inactivated by allopurinol. At infinite concentrations of reducing and oxidizing substrates, the Km for hypoxanthine was 119 microM, for nicotinic acid was 136 microM, and for NAD+ was 525 microM. 相似文献
Large amounts of Cu-metallothionein were obtained by 2-mercaptoethanol and sodium dodecyl sulphate extractions of the particulate fractions of the liver of pigs given high-Cu2+ diets or rats injected with Cu2+. Three isoproteins were purified from pig liver and characterized on the basis of their physicochemical properties, metal content and amino acid composition. No such pool of Cu-metallothionein was present in the liver of Cu2+-loaded sheep or of rats given Cu2+-supplemented diets. 相似文献
Approximately 43–60% of the total genome in bovine, goat and sheep consisted of interspersed repeated and single copy DNA
sequences. Most of the interspersed repeated DNA sequences were 1500–2400 nucleotide pair long while a minor portion was more
than 4000 nucleotide pair long in goat and sheep and 3200 nucleotide pair long in bovine. About 1/3rd of single copy sequence
were interspersed and their length was in the range of 1000–1500 nucleotide pairs. 相似文献
The rapid and aggressive spread of artemisinin-resistant Plasmodium falciparum carrying the C580Y mutation in the kelch13 gene is a growing threat to malaria elimination in Southeast Asia, but there is no evidence of their spread to other regions. We conducted cross-sectional surveys in 2016 and 2017 at two clinics in Wewak, Papua New Guinea (PNG) where we identified three infections caused by C580Y mutants among 239 genotyped clinical samples. One of these mutants exhibited the highest survival rate (6.8%) among all parasites surveyed in ring-stage survival assays (RSA) for artemisinin. Analyses of kelch13 flanking regions, and comparisons of deep sequencing data from 389 clinical samples from PNG, Indonesian Papua and Western Cambodia, suggested an independent origin of the Wewak C580Y mutation, showing that the mutants possess several distinctive genetic features. Identity by descent (IBD) showed that multiple portions of the mutants’ genomes share a common origin with parasites found in Indonesian Papua, comprising several mutations within genes previously associated with drug resistance, such as mdr1, ferredoxin, atg18 and pnp. These findings suggest that a P. falciparum lineage circulating on the island of New Guinea has gradually acquired a complex ensemble of variants, including kelch13 C580Y, which have affected the parasites’ drug sensitivity. This worrying development reinforces the need for increased surveillance of the evolving parasite populations on the island, to contain the spread of resistance. 相似文献
Riparian zones are among the most valuable ecosystems on the earth. They act as the ecological engineers that improve river health through delivering a range of ecosystem functions. Stream bank stabilization, pollutant and sediment buffering, temperature regulation, provision of energy to river food webs and communities, groundwater recharge and provision of ecological corridors and habitat for wildlife, are among major ecosystem functions of riparian zones that play a great role in river health. Besides these ecosystem functions, riparian zones also provide various ecosystem goods and services for human well-being. But in the current scenario, riparian zones are under severe threat due to agricultural activities, urbanization, river flow alteration, overexploitation, climate change, pollution, and biological invasion. In the present and probable future scenarios of declining river health and global environmental changes, there is a pressing need of an integrated approach for managing riparian zones. This review article aims to advocate an integrated approach for riparian zone management based on various components such as riparian condition assessment, policy framework, stakeholder’s participation, management practices, legislation, and awareness. Authors also discussed riparian zones in context of their concepts, features, functions, and threats.