The rust mycobiota of southern Africa: Species richness, composition, and affinities

The rust mycobiota (Uredinales, Basidiomycota) of southern Africa (Botswana, Namibia, and South Africa) is analysed with regard to species richness, generic composition, and similarities to the rust mycobiotas of the remaining African continent and other regions of the world. Southern Africa is home to about 546 rust species: ca 522 species have been reported from South Africa, 73 from Namibia, and less than ten from Botswana. Thirty-two species were considered to be exotics. Two hundred and twenty-five of the species are restricted to southern Africa, suggesting an endemism rate of ca 44 %. At present, the rust fungus:host ratio is 1:38.5, which is much lower than expected from other regions of the world. This low ratio may partly be due to under-exploration of the area, but the results presented here indicate that a natural paucity of rust fungi on certain, especially species-rich plant taxa centred in southern Africa and possibly environmental factors are more important reasons. The predominant genera are Puccinia and Uromyces accounting for ca 59 % of the rust species. The genera Hemileia, Phakopsora and especially Ravenelia, centred in tropical regions, are well represented and sum up to 8 % of the species. Members of Melampsoraceae and Phragmidiaceae, common in temperate regions of the Northern Hemisphere, are scarce. Most of the other 28 recorded teleomorph genera are only represented by three or less species. In an African context, most species are shared with central and east Africa (almost 16 %). Only a few species are disjunct between southern and West Africa or Madagascar. Ca 10 % of the species are shared only with other parts of the paleotropics, especially the Indian subcontinent. Disjunctions of native species with the New World, Australia/New Zealand, or Europe are rare.     

Immunolocalization of MMP-19 in the human intervertebral disc: implications for disc aging and degeneration

Matrix metalloproteinases (MMPs) degrade components of the extracellular matrix of the disc, but the presence of MMP-19 has not been explored. In other tissues, MMP-19 is known to act in proteolysis of the insulin-like growth factor (IGF) binding protein-3, thereby exposing this protein to make it available to influence cell behavior. MMP-19 also has been shown to inhibit capillary-like formation and thus play a role in the avascular nature of the disc. Using immunohistochemistry, normal discs from six subjects aged newborn through 10 years and 20 disc specimens from control donors or surgical patients aged 15-76 (mean age 40.2 years) were examined for immunolocalization of MMP-19; six Thompson grade I discs, five Thompson grade II, eight Thompson grade III, five Thompson grade IV, and one Thompson grade V discs were analyzed. The results indicate that in discs from young subjects, MMP-19 was uniformly localized in the outer annulus. In discs from adult donors and surgical patients, outer and inner annulus cells only occasionally showed MMP-19 localization. The greatest expression of MMP-19 was observed in young discs, and little expression was seen in older or degenerating discs. Because MMP-19 has been shown to regulate IGF-mediated proliferation in other tissues, its decline in the aging/degenerating disc may contribute to the age-related decrease in disc cell numbers.     

Pathological shear stress stimulates the tyrosine phosphorylation of alpha-actinin associated with the glycoprotein Ib-IX complex.

Shear-induced platelet responses are triggered by VWF binding to the platelet GpIb-IX complex, and there is evidence that this ligand-receptor coupling stimulates transmembranous signaling through the cytoplasmic tail of glycoprotein (Gp) Ib alpha. To investigate the mechanism by which signaling is effected, new molecular interactions involving GpIb-IX that develop in response to pathological shearing stress were examined in intact human platelets. Exposure to shear, but not alpha-thrombin, results in the co-immunoprecipitation of the actin cross-linking protein alpha-actinin with the GpIb-IX complex. Blockers of VWF binding to GpIb alpha or actin polymerization inhibit the association of alpha-actinin with the GpIb-IX complex, but the association of alpha-actinin with the GpIb-IX complex is not affected by inhibiting VWF binding to platelet integrin alpha IIb beta 3 (GpIIb-IIIa). alpha-Actinin becomes tyrosine phosphorylated in response to pathological shear stress, and phosphorylated alpha-actinin associates with GpIb-IX. In resting platelets, class IA heterodimeric phosphatidylinositol 3-kinase (PI 3-K) and protein kinase N (PKN) associate with nonphosphorylated alpha-actinin. Shear stress causes PI 3-K to disassociate from alpha-actinin, while it stimulates PKN binding to alpha-actinin. These results demonstrate that shear-induced VWF binding to GpIb alpha causes enhanced binding of cytoskeletal alpha-actinin to GpIb-IX and suggest that alpha-actinin, perhaps through tyrosine phosphorylation, serves as an adapter for a signaling complex that could regulate VWF-induced platelet aggregation.     

Structure, dynamics and electrostatics of the active site of glutaredoxin 3 from Escherichia coli: comparison with functionally related proteins.

The chemistry of active-site cysteine residues is central to the activity of thiol-disulfide oxidoreductases of the thioredoxin superfamily. In these reactions, a nucleophilic thiolate is required, but the associated pK(a) values differ vastly in the superfamily, from less than 4 in DsbA to greater than 7 in Trx. The factors that stabilize this thiolate are, however, not clearly established. The glutaredoxins (Grxs), which are members of this superfamily, contain a Cys-Pro-Tyr-Cys motif in their active site. In reduced Grxs, the pK(a) of the N-terminal active-site nucleophilic cysteine residue is lowered significantly, and the stabilization of the corresponding thiolate is expected to influence the redox potential of these enzymes. Here, we use a combination of long molecular dynamics (MD) simulations, pK(a) calculations, and experimental investigations to derive the structure and dynamics of the reduced active site from Escherichia coli Grx3, and investigate the factors that stabilize the thiolate. Several different MD simulations converged toward a consensus conformation for the active-site cysteine residues (Cys11 and Cys14), after a number of local conformational changes. Key features of the model were tested experimentally by measurement of NMR scalar coupling constants, and determination of pK(a) values of selected residues. The pK(a) values of the Grx3 active-site residues were calculated during the MD simulations, and support the underlying structural model. The structure of Grx3, in combination with the pK(a) calculations, indicate that the pK(a) of the N-terminal active-site cysteine residue in Grx3 is intermediate between that of its counterpart in DsbA and Trx. The pK(a) values in best agreement with experiment are obtained with a low (<4) protein dielectric constant. The calculated pK(a) values fluctuate significantly in response to protein dynamics, which underscores the importance of the details of the underlying structures when calculating pK(a) values. The thiolate of Cys11 is stabilized primarily by direct hydrogen bonding with the amide protons of Tyr13 and Cys14 and the thiol proton of Cys14, rather than by long-range interactions from charged groups or from a helix macrodipole. From the comparison of reduced Grx3 with other members of the thioredoxin superfamily, a unifying theme for the structural basis of thiol pK(a) differences in this superfamily begins to emerge.     

Comparative ecological analysis of Toarcian (Lower Jurassic) benthic faunas from southern France and east-central Spain

Toarcian (Lower Jurassic) argillaceous-marly sediments of southern France (Causses) and east-central Spain (Barranco de la Puta section) were deposited in mid to lower shelf settings. They contain a low to moderately diverse, autochthonous fauna of benthic macroinvertebrates dominated by bivalves and gastropods, in the case of the Causses, and by bivalves and brachiopods in the Barranco de la Puta section. Five benthic associations are recognized at the latter locality and four at the former. In the Causses, low species diversity, evidence of seasonal mass mortality, presence of the opportunistic bivalve Parvamussium pumilum, local dominance of the soft bottom coral Thecocyathus mactrus and the dark-grey, fine-grained sediment point to high rates of sedimentation, high turbidity, intermittently lowered oxygen values and soft to soupy substrate conditions. These factors, together with eutrophic water masses, were the main environmental parameters governing faunal distribution. Soupy substrate conditions are thought to be largely biologically produced by the activity of infaunal depositfeeders, chiefly nuculoid bivalves. In the Barranco de la Puta section, abundant brachiopods, higher carbonate content of the sediment, and evidence of intermittent in-situ reworking and winnowing point to an influence of storm-induced currents, a shallower depositional depth, lower turbidity, a lower rate of sedimentation, and a somewhat firmer substrate. As a consequence, the fauna is largely dominated by epifaunal suspension-feeders. Compared with maximum and average sizes from elsewhere in the Jurassic, most bivalve species are distinctly smaller, a feature interpreted as stunting. Brachiopods, in contrast, reach normal size. The lack of nuculids, scarcity of deposit-feeders in general, and stunting are interpreted as evidence of an oligotrophic environment. Lowered oxygen conditions appear to have played only a very limited role in shaping the distribution pattern. Many of the differences between both areas can be explained by the different trophic regimes that reflect considerably higher run-off and consequently higher input of terrigenous sediment and dissolved nutrients to the basin in the case of the Causses. This in turn is thought to possibly reflect differences in regional climate; humid in southern France and more arid in eastcentral Spain.     

Rat heart glycogen phosphorylase II is genetically distinct from phosphorylase I

Previous studies in our laboratory have shown that rat heart glycogen phosphorylase (1,4-alpha-D-glucan: orthophosphate alpha-D-glucosyltransferase, EC 2.4.1.1) separates into two forms upon ion-exchange chromatography. Both forms could be shown to have the same subunit Mr and to incorporate one molecule of phosphate per subunit. The studies reported here were done to check whether both forms are native isoenzymes and, further, which form might represent the heart-specific phosphorylase. Firstly, the iso-electric points of the purified enzymes are compared with those associated with phosphorylase activity in crude extracts from rat heart. Two out of four major bands coincided with the bands of purified phosphorylase Ib and IIb (isoelectric points: 5.5 and 6.25), indicating apparent identity. Secondly, antibodies to rat skeletal muscle phosphorylase reacted with rat heart phosphorylase I, whereas phosphorylase II was neither inhibited nor precipitated by the antibody. Thirdly, peptide maps obtained after proteolytic digestion of SDS-denatured phosphorylase I and II showed different patterns. In addition to the kinetic differences between these two forms reported earlier, phosphorylase IIa was inhibited by glucose 6-phosphate, whereas phosphorylase Ia was not. These results suggest that phosphorylase II is a heart-specific isoenzyme which is presumably encoded by a different gene.     

Determination of the solution structure of a platelet-adhesion peptide of von Willebrand factor.

Two-dimensional nuclear magnetic resonance (NMR) spectroscopy in combination with distance geometry (DG) and dynamical simulated annealing (DSA) calculations have been used to determine the tertiary solution structure of a synthetic 29-residue fragment of von Willebrand factor (vWF). This fragment (D514-E542) represents an adhesion site on vWF for its platelet receptor, the glycoprotein Ib-IX complex (GP Ib-IX). The NMR data yielded 109 interproton distance measurements and two chi 1 dihedral angle constraints for use in DG and DSA calculations. Most prominent in the calculated family of solution structures was an amphipathic, right-handed alpha-helix in the C-terminal segment of the peptide. We propose that this highly structured region may be important for the specific molecular interaction of vWF with the GP Ib-IX complex.