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Query: UNIPROT:P06889 (Mol)
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The molecular characterization of 14 strains of Kluyveromyces marxianus isolated from Agave fourcroydes (Lem.) in Yucatan, Mexico, was performed by AP-PCR analysis, PCR-RFLP of 5.8S-ITS, and complete NTS regions. A sequence analysis of the D1/D2 domain of the 26S rDNA was also carried out in six selected strains. The AP-PCR approach had the highest discrimination power for the molecular characterization of new henequen K. marxianus strains. PCR-RFLP of 5.8S-ITS regions did not reveal polymorphisms in this group of strains. The restriction enzyme digestion analysis of NTS region enables the separation among strains which coincides with ascospore shape groups. The molecular tools used in this article may be useful to confirm a preliminary screen of yeasts isolated from henequen without the use of growth characteristics or morpho-physiological tests.
Mol Biotechnol 2007 Nov
PMID:Molecular characterization of Kluyveromyces marxianus strains isolated from Agave fourcroydes (Lem.) in Yucatan, Mexico. 1795 62

Symbiotic nitrogen fixation with nitrogen-fixing bacteria in the root nodules is a distinctly beneficial metabolic process in legume plants. Legumes control the nodule number and nodulation zone through a systemic negative regulatory system between shoot and root. Mutation in the soybean NTS gene encoding GmNARK, a CLAVATA1-like serine/threonine receptor-like kinase, causes excessive nodule development called hypernodulation. To examine the effect of nts mutation on the gene expression profile in the leaves, suppression subtractive hybridization was performed with the trifoliate leaves of nts mutant 'SS2-2' and the wild-type (WT) parent Sinpaldalkong2, and 75 EST clones that were highly expressed in the leaves of the SS2-2 mutant were identified. Interestingly, the expression of jasmonate (JA)-responsive genes such as vspA, vspB, and Lox2 were upregulated, whereas that of a salicylate-responsive gene PR1a was suppressed in the SS2-2 mutant. In addition, the level of JA was about two-fold higher in the leaves of the SS2-2 mutant than in those of the WT under natural growth conditions. Moreover, the JA-responsive gene expression persists in the leaves of SS2-2 mutant without rhizobia infection in the roots. Taken together, our results suggest that the nts mutation increases JA synthesis in mature leaves and consequently leads to constitutive expression of JA-responsive genes which is irrelevant to hypernodulation in the root.
Mol Cells 2007 Oct 31
PMID:The Hypernodulating nts mutation induces jasmonate synthetic pathway in soybean leaves. 1797 70

The phylogenetic utility of chloroplast (atpB-rbcL, petD, rps16, trnL-F) and nuclear (ETS, ITS) DNA regions was investigated for the tribe Spermacoceae of the coffee family (Rubiaceae). ITS was, despite often raised cautions of its utility at higher taxonomic levels, shown to provide the highest number of parsimony informative characters, in partitioned Bayesian analyses it yielded the fewest trees in the 95% credible set, it resolved the highest proportion of well resolved clades, and was the most accurate region as measured by the partition metric and the proportion of correctly resolved clades (well supported clades retrieved from a combined analysis regarded as "true"). For Hedyotis, the nuclear 5S-NTS was shown to be potentially as useful as ITS, despite its shorter sequence length. The chloroplast region being the most phylogenetically informative was the petD group II intron. We also present a phylogeny of Spermacoceae based on a Bayesian analysis of the four chloroplast regions, ITS, and ETS combined. Spermacoceae are shown to be monophyletic. Clades supported by high posterior probabilities are discussed, especially in respect to the current generic classification. Notably, Oldenlandia is polyphyletic, the two subgenera of Kohautia are not sister taxa, and Hedyotis should be treated in a narrow sense to include only Asian species.
Mol Phylogenet Evol 2008 Dec
PMID:The phylogenetic utility of chloroplast and nuclear DNA markers and the phylogeny of the Rubiaceae tribe Spermacoceae. 1895 Jul 20

Dysfunction of vascular nitric oxide (NO)/cGMP signaling is believed to contribute essentially to various cardiovascular disorders. Besides synthesis and/or bioavailability of endothelial NO, impaired function of soluble guanylate cyclase (sGC) may play a key role in vascular dysfunction. Based on the proposal that desensitization of sGC through S-nitrosation contributes to vascular NO resistance ( Proc Natl Acad Sci U S A 104: 12312-12317, 2007 ), we exposed purified sGC to dinitrosyl iron complexes (DNICs), known as potent nitrosating agents. In the presence of 2 mM GSH, DNICs stimulated cGMP formation with EC(50) values of 0.1 to 0.5 microM and with an efficacy of 70 to 80% of maximal activity measured with 10 microM 2,2-diethyl-1-nitroso-oxyhydrazine (DEA/NO). In the absence of GSH, the efficacy of DNICs was markedly reduced, and sGC stimulation was counteracted by the inhibition of both basal and DEA/NO-stimulated cGMP formation at higher DNIC concentrations. Inactivation of sGC was slowly reversed in the presence of 2 mM GSH and associated with stoichiometric S-nitrosation of the protein (2.05 +/- 0.18 mol S-nitrosothiol per mol of 143-kDa heterodimer). S-Nitrosoglutathione and sodium nitroprusside caused partial inhibition of DEA/NO-stimulated sGC that was prevented by GSH, whereas nitroglycerin (0.3 mM) had no effect. Our findings indicate that nitrosation of two cysteine residues in sGC heterodimers results in enzyme inactivation. Protection by physiologically relevant concentrations of GSH (10 microM to 3 mM) suggests that S-nitrosation of sGC may contribute to vascular dysfunction in inflammatory disorders associated with nitrosative and oxidative stress and GSH depletion.
Mol Pharmacol 2009 Apr
PMID:Inactivation of soluble guanylate cyclase by stoichiometric S-nitrosation. 1911 87

Adelges cooleyi is a host-alternating, gall-making insect native to the Rocky Mountains and Cascade Mountains in western North America. The insect's primary hosts are Picea (spruce) species, and its secondary host is Pseudotsuga menziesii, Douglas fir. To determine whether there are large-scale patterns of genetic variation in this specialist insect, we created molecular phylogenies of geographically separate samples of A. cooleyi using sequence data from two mitochondrial (mtDNA) genes and amplified fragment length polymorphisms (AFLPs). Three divergent mtDNA lineages were identified. Analysis of mtDNA and AFLP genetic variation revealed that samples from southeastern Arizona are genetically isolated from all other samples. AFLP data identified possible gene flow between individuals from divergent mtDNA lineages in an area in the central Rocky Mountains. Factors that likely affected divergence within A. cooleyi were identified by comparing our conclusions with well-known changes in the distribution of vegetation in response to glaciations and previous phylogeographical work conducted on this specialist insect's host-plants. In addition to documenting previously unknown patterns of genetic variation in A. cooleyi, our work provides the basis for a testable hypothesis regarding the extent to which the distribution of variation in Picea and Pseudotsuga hosts mediates the distribution of genetic variation for this specialist insect.
Mol Ecol 2009 Jan
PMID:Phylogeography of a specialist insect, Adelges cooleyi: historical and contemporary processes shape the distribution of population genetic variation. 1919 84

Mammalian myeloid and epithelial cells express various peptide antibiotics (such as defensin and cathelicidin) that contribute to the innate host defense against invading microorganisms. Among these, guinea pig cathelicidin CAP11 (G1-I43) possesses potent antibacterial activities against Gram-positive and -negative bacteria, and also lipopolysaccharide-neutralizing activity. We previously revealed that the active region with antibacterial activity is localized at G1 to R18 of CAP11. In this study, to develop peptide derivatives with enhanced antimicrobial actions, we utilized the amphipathic 18-mer peptide (G1-R18) as a template. Anti-microbial activities of the peptides were assessed by alamarBlue assay (Escherichia coli, Staphylococcus aureus and Candida albicans) and colony formation assay (Porphyromonas gingivalis). Furthermore, the membrane-permeabilization activities were determined by using E. coli ML-35p as a target. By substituting K5, T9, R10, R12, and G17 with five L residues, the hydrophobicity of the peptide (1-18m1) was increased, and by substituting G1, and Q14 with K and R residues, respectively, the hydrophilicity (positive charge) of the peptide (1-18m2) was enhanced. Among the peptides, 1-18m2 exhibits the most potent antimicrobial and membrane-permeabilizing activities against the microorganisms examined. Thus, the antimicrobial activities of the amphipathic CAP11-derived 18-mer peptide can be augmented by modifying its hydrophobicity and hydrophilicity (positive charge), and 1-18m2 is the most potent among the peptide derivatives with therapeutic potential for Gram-positive and -negative bacterial, and fungal infections.
Int J Mol Med 2009 Apr
PMID:Augmentation of the antimicrobial activities of guinea pig cathelicidin CAP11-derived peptides by amino acid substitutions. 1928 26

Quaternary climatic oscillations greatly influenced the present-day population genetic structure of animals and plants. For species with high dispersal and reproductive potential, phylogeographic patterns resulting from historical processes can be cryptic, overshadowed by contemporary processes. Here we report a study of the phylogeography of Odocoileus hemionus, a large, vagile ungulate common throughout western North America. We examined sequence variation of mitochondrial DNA (control region and cytochrome b) within and among 70 natural populations across the entire range of the species. Among the 1766 individual animals surveyed, we recovered 496 haplotypes. Although fine-scale phylogenetic structure was weakly resolved using phylogenetic methods, network analysis clearly revealed the presence of 12 distinct haplogroups. The spatial distribution of haplogroups showed a strong genetic discontinuity between the two morphological types of O. hemionus, mule deer and black-tailed deer, east and west of the Cascade Mountains in the Pacific Northwest. Within the mule deer lineage, we identified several haplogroups that expanded before or during the Last Glacial Maximum, suggesting that mule deer persisted in multiple refugia south of the ice sheets. Patterns of genetic diversity within the black-tailed deer lineage suggest a single refugium along the Pacific Northwest coast, and refute the hypothesis that black-tailed deer persisted in one or more northern refugia. Our data suggest that black-tailed deer recolonized areas in accordance with the pattern of glacial retreat, with initial recolonization northward along a coastal route and secondary recolonization inland.
Mol Ecol 2009 Apr
PMID:Species-wide phylogeography of North American mule deer (Odocoileus hemionus): cryptic glacial refugia and postglacial recolonization. 1930 64

We aim to develop a cultured cell model, to serve as a system with which the altered circadian phenotypes produced by the clock gene variations could be studied in vitro. Tau mutation, which shortens the circadian period of hamsters and mice, was introduced into the CK1epsilon locus of cultured Rat1-R12 cells by gene targeting mediated by a recombinant adeno-associated virus (rAAV) vector. After transduction of Rat1-R12 cells with rAAV, about 0.14% of the drug-resistant cells underwent gene targeting at CK1epsilon locus. Of the three clones isolated, only one carried the targeted allele of tau mutation and two carried the targeted wild-type allele. The clone with the targeted tau mutant allele exhibited a significantly shorter circadian period compared to the clone with targeted wild-type allele. rAAV-mediated gene targeting in cultured somatic cells is a convenient and powerful tool for analyzing the phenotypic outcome of clock gene variations, and for elucidating the pathogenesis of the disorders associated with abnormal circadian rhythmicity.
Cell Mol Neurobiol 2009 Jul
PMID:Introduction of tau mutation into cultured Rat1-R12 cells by gene targeting, using recombinant adeno-associated virus vector. 1930 50

Nitroxyl (HNO) may be formed endogenously by uncoupled nitric-oxide (NO) synthases, enzymatic reduction of NO or as product of vascular nitroglycerin bioactivation. The established HNO donor Angeli's salt (trioxodinitrate, AS) causes cGMP-dependent vasodilation through activation of soluble guanylate cyclase (sGC). We investigated the mechanisms underlying this effect using purified sGC and cultured endothelial cells. AS (up to 0.1 mM) had no significant effect on sGC activity in the absence of superoxide dismutase (SOD) or dithiothreitol (DTT). In the presence of SOD, AS caused biphasic sGC activation (apparent EC(50) approximately 10 nM, maximum at 1 microM) that was accompanied by the formation of NO. DTT (2 mM) inhibited the effects of <10 microM AS but led to sGC activation and NO release at 0.1 mM AS even without SOD. AS had no effect on ferric sGC, excluding activation of the oxidized enzyme by HNO. The NO scavenger carboxy-PTIO inhibited endothelial cGMP accumulation induced by AS in the presence but not in the absence of SOD (EC(50) approximately 50 nM and approximately 16 microM, respectively). Carboxy-PTIO (0.1 mM) inhibited the effect of <or=10 microM AS in the presence of SOD but caused NO release from 0.1 mM AS in the absence of SOD. These data indicate that AS activates sGC exclusively via NO, formed either via SOD-catalyzed oxidation of HNO or through a minor AS decomposition pathway that is unmasked in the presence of HNO scavenging thiols.
Mol Pharmacol 2009 Nov
PMID:Mechanisms underlying activation of soluble guanylate cyclase by the nitroxyl donor Angeli's salt. 1972 Jul 27

Recent human activities have spread numerous plant species across the globe, yet it is unclear to what degree historical human activities influenced plant dispersal. In western North America, Camassia quamash was one of the most important food plants for indigenous peoples, who transported its propagules either intentionally or accidentally. We investigated how human and natural dispersal might have contributed to the current pattern of spatial genetic structure in C. quamash by performing phylogeographical surveys at two geographical scales. We sequenced two noncoding regions of chloroplast deoxyribonucleic acid (DNA) in 226 individuals from 53 populations of C. quamash as well as 126 individuals from 21 populations of the non-food plant Zigadenus venenosus. Contrary to the expectation of anthropogenic transport, C. quamash populations did not exhibit weaker genetic structure than Z. venenosus populations. We also failed to find convincing evidence for signatures of transport. Instead, our data showed strong effects of past glaciation and geographical barriers of the mountains in the Cascade Range, Olympic Peninsula and Vancouver Island. West of the Cascades, the species appears to have largely migrated northward from a southern refugium after deglaciation, whereas few populations having a highly divergent haplotype might have survived in southwestern Washington. Our data suggest that despite substantial ethnobotanical evidence for anthropogenic transport, the current pattern of genetic structure of C. quamash does not show any detectable signatures of transport by indigenous peoples and is better understood as the result of natural dispersal processes.
Mol Ecol 2009 Sep
PMID:Phylogeography of Camassia quamash in western North America: postglacial colonization and transport by indigenous peoples. 1973 32


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