B-ring of colchicine and its role in taxol-induced tubulin polymerization

Taxol-induced assembly of purified tubulin is not inhibited by the colchicine analogue 2-methoxy-5-(2',3',4'-trimethoxyphenyl)tropone. Colchicine analogues having intact A, C and B-rings (without NH-CO-CH3) such as desacetamidocolchicine have also been found to be inactive. It has been observed that these two colchicine analogues are incorporated into polymers when incubated in the presence of taxol. Furthermore, preformed taxol-induced polymers of tubulin have been found to bind these two colchicine analogues. These results suggest that colchicine-binding domains on the tubulin molecule are mostly (if not completely) exposed in the taxol-induced polymers.     

Nitrogen and carbon mineralization during incubation of two Bangladesh soils in relation to temperature

Summary At temperatures of 20°, 30°, 40°, 50° and 60°C in a Gangetic alluvial soil (G soil, pH 7.6) N-mineralization and nitrification increased with temperature up to 40°C and mineralized N accumulated entirely as nitrate. At 50° and 60°C mineralized N was relatively low and no nitrification occurred. In the Red soil (R soil, pH 5.2) mineralized N increased with temperature up to 40°C, was somewhat less at 50°C and was at a maximum at 60°C. Nitrification was maximum at 30°C but did not occur at 50° and 60°C. In the G soil C-mineralization increased considerably with temperature, whilst in the R soil there were only small differences due to temperature.     

Reducing sugar accumulation from alkali pretreated sugar cane bagasse using Cellulomonas

Summary An active cellulolytic culture was obtained following growth of the Cellulomonas strain CS1-17 for 24 h at 32° C on 1 or 2% alkali pretreated sugar cane bagasse. Environmental conditions were then varied to favour reducing sugar accumulation from fresh alkali pretreated bagasse added to the 24 h culture medium at 75 g/l. After incubation for an additional 48 h at 37° C under anaerobic, aerobic and aerobic+0.2% sodium azide conditions, reducing sugar was accumulated at 22.8, 23.7 and 25.6 g/l respectively. Approximately 83% of this release occurred during the first 18 h of incubation and the reducing sugar released contained approximately 14% xylose, 35% glucose, and 26% cellobiose. Addition of exogenous cellobiase resulted in conversion of the cellobiose to glucose.     

Systematics of the Deropristiidae Cable & Hunninen, 1942 (Trematoda) and biogeographical associations with sturgeons (Osteichthyes: Acipenseridae)

The 11 nominal species of the Deropristiidae, belonging to the genera Deropristis (three species), Pristicola (one species), Skrjabinopsolus (five species) and Cestrahelmins (two species) were re-evaluated for a phylogenetic and biogeographical analysis of the group. Skrjabinopsolus elongatus (Madhavi, 1974) (= S. indicus Gupta & Ahmad, 1976, S. kurotchkini Parukhin, 1976), S. sanyaensis Shen, 1990, Deropristis paurosoma Wang, 1989 and Cestrahelmins Fischthal, 1957, do not belong to the Deropristiidae. The genus Opisthodiplomonorchis Madhavi, 1974 is resurrected for its type-species, O. elongatus (= S. elongatus), and D. paurosoma is also transferred to this genus. Species of Opisthodiplomonorchis occur in polynemid fishes of the Indo-Pacific. Five nominal species thus comprise the Deropristiidae, i.e. S. armatus Ivanov in Ivanov and Mrygin, 1937, S. manteri (Cable, 1952), D. hispida (Abildgaard, 1819), D. inflata (Molin, 1859) and P. sturionis (Little, 1930), which corresponds to the original composition of the family. A comparison of Skrjabinopsolus from acipenserids in North America and Europe revealed variability in morphological characters considered diagnostic in the past (body size and position of the ovary and testes). Based on the morphology and biology, the two species of Skrjabinopsolus can be considered incipient species. A phylogenetic analysis of the redefined Deropristiidae indicates that Deropristis and Pristicola may form a monophyletic group with Skrjabinopsolus as the sister taxon. The biogeography of the Deropristiidae suggests an origin and distribution associated with the formation of the North Atlantic basin. Freshwater and marine/estuarine origins for the family are equally parsimonious possibilities. A freshwater origin can be traced at least to Cretaceous Laurasia followed by a drift-vicariance separation and subsequent association with North Atlantic drainages. Support for a marine ancestry of the Deropristiidae is indicated by the biogeography of its sister taxa (outgroups in this study) and of its hosts, and is also consistent with an Atlantic origin. The deropristiids evidently originated and speciated within the Acipenseridae in the Atlantic region, with a host switch into Atlantic Anguilla spp.     

Oligosaccharide modification in the early secretory pathway directs the selection of a misfolded glycoprotein for degradation by the proteasome

The role of conformation-based quality control in the early secretory pathway is to eliminate misfolded polypeptides and unassembled multimeric protein complexes from the endoplasmic reticulum, ensuring the deployment of only functional molecules to distal sites. The intracellular fate of terminally misfolded human alpha1-antitrypsin was examined in hepatoma cells to identify the functional role of asparagine-linked oligosaccharide modification in the selection of glycoproteins for degradation by the cytosolic proteasome. Proteasomal degradation required physical interaction with the molecular chaperone calnexin. Altered sedimentation of intracellular complexes following treatment with the specific proteasome inhibitor lactacystin, and in combination with mannosidase inhibition, revealed that the removal of mannose from attached oligosaccharides abrogates the release of misfolded alpha1-antitrypsin from calnexin prior to proteasomal degradation. Intracellular turnover was arrested with kifunensine, implicating the participation of endoplasmic reticulum mannosidase I in the disposal process. Accelerated degradation occurred in a mannosidase-independent manner and was arrested by lactacystin, in response to the posttranslational inhibition of glucosidase II, demonstrating that the attenuated removal of glucose from attached oligosaccharides functions as the underlying rate-limiting step in the proteasome-mediated pathway. A model is proposed in which the removal of mannose from multiple attached oligosaccharides directs calnexin in the selection of misfolded alpha1-antitrypsin for degradation by the proteasome.     

Examination of the nickel site structure and reaction mechanism in Streptomyces seoulensis superoxide dismutase

Superoxide dismutases are metalloenzymes involved in protecting cells from oxidative damage arising from superoxide radical or reactive oxygen species produced from superoxide. Examples of enzymes containing Cu, Mn, and Fe as the redox-active metal have been characterized. Recently, a SOD containing one Ni atom per subunit was reported. The amino acid sequence of the NiSOD deduced from the nucleotide sequence of the structural gene sodN from Streptomyces seoulensis is reported and has no homology with other SODs. X-ray absorption spectroscopic studies coupled with EPR of the Ni center show that the Ni in the oxidized (as isolated) enzyme is in a five-coordinate site composed of three S-donor ligands, one N-donor, and one other O- or N-donor. This unique coordination environment is modified by the loss of one N- (or O-) donor ligand in the dithionite-reduced enzyme. The NiSOD activity was determined by pulse radiolysis, and a value of kcat = 1.3 x 10(9) M-1 s-1 per Ni was obtained. The rate is pH sensitive and drops off rapidly above pH 8. The results characterize a novel class of metal center active in catalyzing the redox chemistry of superoxide and, when placed in context with other nickel enzymes, suggest that thiolate ligation is a prerequisite for redox-active nickel sites in metalloenzymes.     

Parasites of lake sturgeon, Acipenser fulvescens (Chondrostei: Acipenseridae), from central Canada

Nineteen species of parasites were recovered from lake sturgeon, Acipenser fulvescens, from four major waterways of Central Canada; the Saskatchewan, Nelson, Winnipeg and Rainy River systems, Twelve of these are new host records. Host specific parasites, Crepidostomum auriculatum , Diclybothrium armatum , Spinitectus acipenseri and Truttaedacnitis clitellarius , forming the core parasite species, were recovered with the highest prevalence (≥70%) and were most widely distributed. Polypodium hydriforme was recovered from only stage IV sturgeon oocytes. With the exception of Pomphorhynchus bulbocolli , prevalence and intensity of endohelminth infections were not correlated with sex or age of host but the distribution of non host-specific parasites among sampling sites was determined by the type and relative abundance of food items consumed. The parasites of lake sturgeon are closely correlated to its diet and the species of parasites recovered are more similar to those of freshwater sturgeon in Eurasia than to other species of Acipenser in North America. The present parasite community of lake sturgeon appears to have been shaped by three major factors; the presence of core parasite species which predates geographic isolation, a benthic freshwater diet which has reshaped the parasite community to one comprising freshwater species and a long association with freshwater habitats which is reflected in the reproductive isolation of the lake sturgeon and lastly, the establishment of a host-specific parasite, Spinitectus acipenseri .     

The Integrity of the HMR complex is necessary for centromeric binding and reproductive isolation in Drosophila

Postzygotic isolation by genomic conflict is a major cause for the formation of species. Despite its importance, the molecular mechanisms that result in the lethality of interspecies hybrids are still largely unclear. The genus Drosophila, which contains over 1600 different species, is one of the best characterized model systems to study these questions. We showed in the past that the expression levels of the two hybrid incompatibility factors Hmr and Lhr diverged in the two closely related Drosophila species, D. melanogaster and D. simulans, resulting in an increased level of both proteins in interspecies hybrids. The overexpression of the two proteins also leads to mitotic defects, a misregulation in the expression of transposable elements and decreased fertility in pure species. In this work, we describe a distinct six subunit protein complex containing HMR and LHR and analyse the effect of Hmr mutations on complex integrity and function. Our experiments suggest that HMR needs to bring together components of centromeric and pericentromeric chromatin to fulfil its physiological function and to cause hybrid male lethality.     

Unusual codon usage bias in low expression genes of Vibrio cholerae

Positive correlation between gene expression and synonymous codon usage bias is well documented in the literature. However, in the present study of Vibrio cholerae genome, we have identified a group of genes having unusually high codon usage bias despite being low potential expressivity. Our results suggest that codon usage in lowly expressed genes might also be selected on to preferably use non-optimal codons to maintain a low cellular concentration of the proteins that they encode. This would predict that lowly expressed genes are also biased in codon usage, but in a way that is opposite to the bias of highly expressed genes.     

Analysis of processivity of mungbean dideoxynucleotide-sensitive DNA polymerase and detection of the activity and expression of the enzyme in the meristematic and meiotic tissues and following DNA damaging agent

Analysis of the processivity of mungbean ddNTP-sensitive DNA polymerase showed the incorporation of ∼35-40 nucleotides per binding event in the replication assays involving M13 ss DNA template with 5′-labeled 17-mer primer. Optimal processivity was obtained with 100-150 mM KCl and 6-8 mM Mg²⁺ at pH 7.5. The enzyme showed preference for Mg²⁺ over Mn²⁺ as the metal activator for processivity. 2′, 3′ dideoxythymidine 5′ triphosphate (ddTTP) and rat DNA pol β antibody strongly influenced distributive synthesis. Considerable enhancement in processivity was noticed at 1 mM ATP and 2-4 mM spermidine while higher concentrations of spermidine caused distributive synthesis. The enzyme was found to be active in both meristematic and meiotic tissues and distinctly induced by EMS treatment. DNA-binding assays revealed distinct binding ability of the enzyme to template/primer and damaged DNA substrate. Together these observations illustrate the probable involvement of the enzyme in replication and repair machinery in higher plants.