Sequence homology between the tyrosine-sensitive 3-deoxy-D-arabino-heptulosonate 7-phosphate synthase from Escherichia coli and hemerythrin from Sipunculida

The first enzyme of the common aromatic biosynthetic pathway in Escherichia coli, the 3-deoxy-D-arabino-heptulosonate 7-phosphate synthase, contains iron as an integral part of the polypeptide chain, and the enzyme shows an absorption maximum around 350 nm (McCandliss, R.J., and Herrmann, K.M. (1978) Proc. Natl. Acad. Sci. U. S. A. 75, 4810-4813). These two properties are also found in hemerythrin, the oxygen carrier of certain marine invertebrates. The amino acid sequence of residues 10 to 18 of the enzyme from E. coli, His-Ile-Thr-Asp-Glu-Gln-Val-Leu-Met, is highly homologous to the sequence of residues 54 to 62 of hemerythrin from Phascolopsis gouldii, His-Phe-Leu-Asn-Glu-Gln-Val-Leu-Met. His54 and Glu58 of hemerythrin have previously been identified through x-ray and protein sequence analysis as iron ligands. We suggest that residues 10 to 18 of the E. coli enzyme represent part of the iron binding fold in this protein, and that His10 and Glu14 are iron ligands.     

Larvalentwicklung und Metamorphose vonPhoronis psammophila (Phoronida,Tentaculata)

The larval development ofPhoronis psammophila Cori is divided into 6 phases (on the basis of increasing pairs of larval tentacles); furthermore an initial and a ripe phase are distinguished. Specific aspects of the development are described: Formation and structure of larval tentacles; anlage of adult tentacles as a thickening in the larval tentacle base; late development of the metasome (larva with 4–6 tentacles); formation of the metasome pouch in the larva with 8 tentacles; enlargement of the apical plate; differentiation of the gut; differentiation of larval nephridia; formation of pigment particles in the larva with 6 tentacles (storage function of pigments and its significance for larval identification); different types of discoflagella in various regions of the body. The larval development shows the following tendencies: Improvement of locomotion; intensification of food filtration; anlage of adult organs in the larva leading to a shortening of metamorphosis duration. The larva ofP. psammophila is compared with those ofP. pallida, P. hippocrepia, andP. vancouverensis. Earlier larval determinations ofP. psammophila (e.g.Actinotrocha sabatieri, A. hatschekii) are shown to have been mistakes. Termination of the postembryonic phase (metamorphosis) can be induced experimentally by bacteria and also by cations. Pure or mixed bacteria cultures must be present at the beginning exponential growth phase. The bacteria density required is 20–94×10⁶ bact.ml^?1 for pure cultures and on the average 28×10⁶ bact. ml^?1 for mixed cultures. Metamorphosis initiation by cations can be induced with CsCl (0.06 M) and RbCl (0.035 M). Metamorphosis ofP. psammophila occurs in 6 phases: larva, ready for metamorphosis; larva, activated by bacteria or ions; evagination of the metasome diverticle, dislocation of gut; losing and swallowing of episphaere and larval tentacles; formation of the youngP. psammophila. All developmental phases are described and compared with those ofP. muelleri; imperfect metamorphosis is characterized and the youngP. psammophila compared with older stages and the adult Phoronis.     

Yeast dipeptidase: active site mapping by kinetic studies with substrates and substrate analogs.

In order to characterize the active site of yeast dipeptidase in more detail, kinetic studies with a variety of dipeptide substrates and substrate analogs were performed. To analyze kinetic data, computer programs were developed which first calculate initial velocities from progress curves and then evaluate the kinetic parameters by nonlinear regression analysis. A free carboxyl group is a prerequisite for binding of dipeptidase substrates; its position relative to the peptide bond must not deviate from the normal L-dipeptide conformation. The spatial arrangement of the terminal ammonium ion seems to be less crucial. The enzyme's substrate specificity clearly reflects the interactions of the substrate amino acid side chains with complementary dipeptidase subsites. The domain of the enzyme in contact with the C-terminal substrate side chain seems to be an open structure of moderately hydrophobic character. In contrast, the binding site for the amino-terminal side chain is a more strongly hydrophobic "pocket" of limited dimensions. The kinetics of inhibition by free amino acids points to an ordered release of products from the enzyme.     

Flavonoid data and populational observations in support of hybrid status for Populus acuminata

Flavonoid distribution and populational composition at the collection sites of several samples of Populus acuminata strongly suggest that this taxon is an inter-sectional hybrid between P. angustifolia, sect. Tacamahaca and any one of two or three different species of the section Aigeiros, when the entire range is considered. In the vicinity of the type locality, the Aigeiros parent is P. sargentii. Within any population, foliage of trees of P. acuminata is characterized by morphological intermediacy and an essentially additive flavonoid profile, as compared to the two parental taxa present. In habit the trees resemble those of P. sargentii, and habitats at the collection sites were found to be somewhat intermediate. Fifteen flavonoid compounds were consistently present in samples of the putative hybrid. Twelve of these had diagnostic value because they were absent in one of the parental species. One compound, apigenin 7-O-diglucoside, was found only in P. acuminata. In part, our identifications correspond to compounds reported by Crawford in a morphological and chemical study of P. acuminiata.     

Intermediate Filaments Play a Pivotal Role in Regulating Cell Architecture and Function

Intermediate filaments (IFs) are composed of one or more members of a large family of cytoskeletal proteins, whose expression is cell- and tissue type-specific. Their importance in regulating the physiological properties of cells is becoming widely recognized in functions ranging from cell motility to signal transduction. IF proteins assemble into nanoscale biopolymers with unique strain-hardening properties that are related to their roles in regulating the mechanical integrity of cells. Furthermore, mutations in the genes encoding IF proteins cause a wide range of human diseases. Due to the number of different types of IF proteins, we have limited this short review to cover structure and function topics mainly related to the simpler homopolymeric IF networks composed of vimentin, and specifically for diseases, the related muscle-specific desmin IF networks.     

UV irradiation inhibits ABC transporters via generation of ADP-ribose by concerted action of poly(ADP-ribose) polymerase-1 and glycohydrolase

ATP-binding cassette (ABC) transporters are involved in the transport of multiple substrates across cellular membranes, including metabolites, proteins, and drugs. Employing a functional fluorochrome export assay, we found that UVB irradiation strongly inhibits the activity of ABC transporters. Specific inhibitors of poly(ADP-ribose) polymerase-1 (PARP-1) restored the function of ABC transporters in UVB-irradiated cells, and PARP-1-deficient cells did not undergo UVB-induced membrane transport inhibition. These data suggest that PARP-1 activation is necessary for ABC transporter functional downregulation. The hydrolysis of poly(ADP-ribose) by poly(ADP-ribose) glycohydrolase (PARG) was also required, since specific PARG inhibitors, which limit the production of ADP-ribose molecules, restored the function of ABC transporters. Furthermore, ADP-ribose molecules potently inhibited the activity of the ABC transporter P-glycoprotein. Hence, poly(ADP-ribose) metabolism appears to play a novel role in the regulation of ABC transporters.     

Retinoic acid differentiation of HL-60 cells promotes cytoskeletal polarization

Retinoic acid (RA) treatment of HL-60 cells in vitro induces granulocytic differentiation, involving reorganization of the nucleus and cytoplasm, development of chemoattractant-directed migration, and eventual apoptosis. The present studies with HL-60/S4 cells document that major elements of the cytoskeleton are changed: actin increases by 50%; vimentin decreases by more than 95%. The cellular content of alpha-tubulin does not significantly change; but the centrosomal-microtubule (MT) array moves away from the lobulating nucleus. Cytoskeletal-modifying chemicals modulate this polarized reorganization: Taxol and cytochalasin D enhance centrosome movement; nocodazole reverses it. Cytoskeletal-modifying chemicals do not appear to affect nuclear lobulation or the integrity of envelope-limited chromatin sheets (ELCS). Employing bcl-2-overexpressing HL-60 cells permitted demonstration of nuclear lobulation, ELCS formation, and centrosome-MT movement concomitantly during RA-induced differentiation, implying independence between the cellular reorganization and apoptotic programs. RA appears to promote an inherent potential in HL-60 cells for cytoskeletal polarization, likely to be important for chemoattractant-directed cell migration, an established characteristic of mature granulocytes.     

Enhancement of viral fusion by nonadsorbing polymers.

Nonadsorbing polymers such as dextran and poly(ethylene glycol) enhance binding as well as extents of fusion of influenza virus with erythrocytes. Kinetics and extent of viral membrane fusion were measured using an assay based on lipid mixing of a fluorescent dye. The effects of nonadsorbing polymers were in the concentration range from 0 to 10 wt%, far below the concentration required to overcome hydration repulsion forces. The enhancing effects were dependent on the molecular weight of nonadsorbing polymer, and only occurred at molecular weight > 1500; this links the phenomena we observe to the so-called "excluded volume effect" of nonadsorbing polymers. The time delay between triggering and the onset of influenza virus fusion was significantly reduced in the presence of nonadsorbing polymers. High molecular weight poly(ethylene glycol) also induced fusion of vesicular stomatitis virus with intact erythrocytes, which do not serve as target of vesicular stomatitis virus fusion in the absence of the polymer. The forces between membranes which determine rate-limiting processes in viral fusion and how they are affected by nonadsorbing polymers are discussed.