Chemical Evolution of Calc-alkaline Magmas during the Ascent through Continental Crust: Constraints from Methana, Aegean Arc

M1 - egaa036Quaternary calc-alkaline andesitic to dacitic lavas effusively erupted on top of about 30 km thick accreted continental crust at Methana peninsula in the western Aegean arc. We present new data of major and trace element concentrations as well as of Sr-Nd-Pb isotope ratios along with mineral compositions of Methana lavas and their mafic enclaves. The enclaves imply a parental basaltic magma and fractional crystallization processes with relatively little crustal assimilation in the deep part of the Methana magma system. The composition of amphibole in some mafic enclaves and lavas indicates deeper crystallization at similar to 25km depth close to the Moho compared with the evolved lavas that formed atPeer reviewe

Effects of the Hydrous Domain in the Mantle Wedge on Magma Formation and Mixing at the Northeast Lau Spreading Center, SW Pacific

Abundant volcanic activity occurs in the back-arc region of the northern Tofua island arc where the Northeast Lau Spreading Center (NELSC) propagates southwards into older crust causing the formation of numerous seamounts at the propagating rift tip. An off-axis volcanic diagonal ridge (DR) occurs at the eastern flank of the NELSC, linking the large rear-arc volcano Niuatahi with the NELSC. New geochemical data from the NELSC, the southern propagator seamounts, and DR reveal that the NELSC lavas are tholeiitic basalts whereas the rear-arc volcanoes typically erupt lavas with boninitic composition. The sharp geochemical boundary probably reflects the viscosity contrast between off-axis hydrous harzburgitic mantle and dry fertile mantle beneath the NELSC. The new data do not indicate an inflow of Samoa plume mantle into the NELSC, confirming previously published He isotope data. The NELSC magmas form by mixing of an enriched and a depleted Indian Ocean-type upper mantle end-member implying a highly heterogeneous upper mantle composition in this area. Most NELSC lavas are little affected by a slab component implying that melting is adiabatic beneath the spreading center. The DR lavas show the influence of a component from the subducted Louisville Seamount Chain, which was previously thought to be restricted to the nearby arc volcanoes Niuatoputapu and Tafahi. This signature is rarely detected along the NELSC implying little mixing of melts from the low-viscosity hydrous portion of the mantle wedge beneath the rear-arc volcanoes into the melting region of the dry mantle beneath the NELSC.Peer reviewe

Vitamin B6 Is Required for Full Motility and Virulence in Helicobacter pylori

Despite recent advances in our understanding of how Helicobacter pylori causes disease, the factors that allow this pathogen to persist in the stomach have not yet been fully characterized. To identify new virulence factors in H. pylori, we generated low-infectivity variants of a mouse-colonizing H. pylori strain using the classical technique of in vitro attenuation. The resulting variants and their highly infectious progenitor bacteria were then analyzed by global gene expression profiling. The gene expression levels of five open reading frames (ORFs) were significantly reduced in low-infectivity variants, with the most significant changes observed for ORFs HP1583 and HP1582. These ORFs were annotated as encoding homologs of the Escherichia coli vitamin B6 biosynthesis enzymes PdxA and PdxJ. Functional complementation studies with E. coli confirmed H. pylori PdxA and PdxJ to be bona fide homologs of vitamin B6 biosynthesis enzymes. Importantly, H. pylori PdxA was required for optimal growth in vitro and was shown to be essential for chronic colonization in mice. In addition to having a well-known metabolic role, vitamin B6 is necessary for the synthesis of glycosylated flagella and for flagellum-based motility in H. pylori. Thus, for the first time, we identify vitamin B6 biosynthesis enzymes as novel virulence factors in bacteria. Interestingly, pdxA and pdxJ orthologs are present in a number of human pathogens, but not in mammalian cells. We therefore propose that PdxA/J enzymes may represent ideal candidates for therapeutic targets against bacterial pathogens

Protein Glycosylation in Helicobacter pylori: Beyond the Flagellins?

Glycosylation of flagellins by pseudaminic acid is required for virulence in Helicobacter pylori. We demonstrate that, in H. pylori, glycosylation extends to proteins other than flagellins and to sugars other than pseudaminic acid. Several candidate glycoproteins distinct from the flagellins were detected via ProQ-emerald staining and DIG- or biotin- hydrazide labeling of the soluble and outer membrane fractions of wild-type H. pylori, suggesting that protein glycosylation is not limited to the flagellins. DIG-hydrazide labeling of proteins from pseudaminic acid biosynthesis pathway mutants showed that the glycosylation of some glycoproteins is not dependent on the pseudaminic acid glycosylation pathway, indicating the existence of a novel glycosylation pathway. Fractions enriched in glycoprotein candidates by ion exchange chromatography were used to extract the sugars by acid hydrolysis. High performance anion exchange chromatography with pulsed amperometric detection revealed characteristic monosaccharide peaks in these extracts. The monosaccharides were then identified by LC-ESI-MS/MS. The spectra are consistent with sugars such as 5,7-diacetamido-3,5,7,9-tetradeoxy-L-glycero-L-manno-nonulosonic acid (Pse5Ac7Ac) previously described on flagellins, 5-acetamidino-7-acetamido-3,5,7,9-tetradeoxy-L-glycero-L-manno-nonulosonic acid (Pse5Am7Ac), bacillosamine derivatives and a potential legionaminic acid derivative (Leg5AmNMe7Ac) which were not previously identified in H. pylori. These data open the way to the study of the mechanism and role of protein glycosylation on protein function and virulence in H. pylori

Modification of the Campylobacter jejuni flagellin glycan by the product of the Cj1295 homopolymeric-tract-containing gene

The Campylobacter jejuni flagellin protein is O-glycosylated with structural analogues of the nine-carbon sugar pseudaminic acid. The most common modifications in the C. jejuni 81-176 strain are the 5,7-di-N-acetylated derivative (Pse5Ac7Ac) and an acetamidino-substituted version (Pse5Am7Ac). Other structures detected include O-acetylated and N-acetylglutamine-substituted derivatives (Pse5Am7Ac8OAc and Pse5Am7Ac8GlnNAc, respectively). Recently, a derivative of pseudaminic acid modified with a di-O-methylglyceroyl group was detected in C. jejuni NCTC 11168 strain. The gene products required for Pse5Ac7Ac biosynthesis have been characterized, but those genes involved in generating other structures have not. We have demonstrated that the mobility of the NCTC 11168 flagellin protein in SDS-PAGE gels can vary spontaneously and we investigated the role of single nucleotide repeats or homopolymeric- tractcontaining genes from the flagellin glycosylation locus in this process. One such gene, Cj1295, was shown to be responsible for structural changes in the flagellin glycoprot ein. Mass spectrometry demonstrated that the Cj1295 gene is required for glycosylation with the di-O-methylglyceroyl-modified version of pseudaminic acid. © 2010 SGM

Neisseria gonorrhoeae O-linked pilin glycosylation: functional analyses define both the biosynthetic pathway and glycan structure

Neisseria gonorrhoeae expresses an O-linked protein glycosylation pathway that targets PilE, the major pilin subunit protein of the Type IV pilus colonization factor. Efforts to define glycan structure and thus the functions of pilin glycosylation (Pgl) components at the molecular level have been hindered by the lack of sensitive methodologies. Here, we utilized a ‘top-down’ mass spectrometric approach to characterize glycan status using intact pilin protein from isogenic mutants. These structural data enabled us to directly infer the function of six components required for pilin glycosylation and to define the glycan repertoire of strain N400. Additionally, we found that the N. gonorrhoeae pilin glycan is O-acetylated, and identified an enzyme essential for this unique modification. We also identified the N. gonorrhoeae pilin oligosaccharyltransferase using bioinformatics and confirmed its role in pilin glycosylation by directed mutagenesis. Finally, we examined the effects of expressing the PglA glycosyltransferase from the Campylobacter jejuni N-linked glycosylation system that adds N-acetylgalactosamine onto undecaprenylpyrophosphate-linked bacillosamine. The results indicate that the C. jejuni and N. gonorrhoeae pathways can interact in the synthesis of O-linked di- and trisaccharides, and therefore provide the first experimental evidence that biosynthesis of the N. gonorrhoeae pilin glycan involves a lipid-linked oligosaccharide precursor. Together, these findings underpin more detailed studies of pilin glycosylation biology in both N. gonorrhoeae and N. meningitidis, and demonstrate how components of bacterial O- and N-linked pathways can be combined in novel glycoengineering strategies

Engineered biosynthetic pathway for sialic acid analogs and its use for the development of anti-infectives

There is clearly an unmet need for anti-influenza drugs that are more effective than those currently available, and ideally that are effective against various strains of influenza and that are not prone to the development of resistance. The unique features of the NRC-IBS technology, with its anticipated efficacy against numerous strains, possibly including avian flu, will enable it to compete effectively against such products. We also note that the market is quite large, and is likely to continue to grow over the coming years. The technology may also result in the development of agents against other viral infections, and against bacterial infections.Peer reviewed: YesNRC publication: Ye

Elucidation of the CMP-pseudaminic acid pathway in Helicobacter pylori: synthesis from UDP-N-acetylglucosamine by a single enzymatic reaction.

NRC publication: Ye

An ExeAB complex in the type II secretion pathway of Aeromonas hydrophilia : effect of ATP-binding cassette mutations on complex formation and function.: Mol.Microbiol.

NRC publication: Ye