Protein synthesis in rabbit reticulocytes XXII+: a heat stable dialyzable factor (EIF-I*) modulates Met-tRNAf binding to EIF-1.
Explore the source record for details and available documents.
Biomedical subjects
Publications and source records attributed to R Roy.
Explore the source record for details and available documents.
A ribosomal salt (0.5 M KCl) wash factor (RF) that reverses inhibition of protein synthesis in heme-deficient reticulocyte lysates has been resolved from the bulk of Met-tRNAfMet-binding factor (EIF-1), Co-EIF-1, and EIF-2 (ternary complex dissociation factor, TDF). The purified RF restores protein synthesis activity of heme-deficient lysates to the level observed in the presence of hemin. No direct correlation exists between amount of EIF-1 activity and ability to reverse inhibition of protein synthesis in heme-deficient lysates. Homogeneous preparations of EIF-1 are completely inactive in reversal of protein synthesis inhibition in heme-deficient lysates. These findings suggest that RF activity is not due to EIF-1 alone but may or may not require EIF-1 as a component of a complex factor.
Protein synthesis in reticulocytes and their lysates is regulated by heme. In heme deficiency a heme-regulated translational inhibitor (HRI) that blocks initiation of polypeptide chains is activated. HRI is a protein kinase (ATP: protein phosphotransferase, EC 2.7.1.37) that specifically phosphorylates the 38,000-dalton subunit of the Met-tRNA(f) (Met) binding factor (IF), which forms a ternary complex with Met-tRNA(f) (Met) and GTP, a finding that suggests that the inhibition by HRI involves the phosphorylation of IF. We have investigated the effect of HRI in the partial reactions of protein chain initiation in which the IF-promoted binding of Met-tRNA(f) (Met) to 40S ribosomal subunits is enhanced by another initiation factor [ternary complex dissociation factor (TDF)] and AUG. The results show that HRI at very low concentrations markedly inhibits the binding of Met-tRNA(f) (Met) to 40S subunits. The inhibitory effect of HRI requires ATP. Under these conditions HRI phosphorylates only the 38,000-dalton subunit of IF. The TDF preparations not only promote the binding of the ternary complex to 40S subunits but also promote the dissociation of the ternary complex in the presence of 5 mM Mg(2+) at 0 degrees . The preincubation of purified IF alone with low concentrations of HRI and ATP does not significantly affect its capacity to form the ternary complex; however, the TDF-promoted dissociation of the ternary complex is inhibited. The nonhydrolyzable analog adenosine 5'-[beta,gamma-imido]triphosphate does not substitute for ATP. These findings suggest that phosphorylation causes a conformational modification in IF, which results in inhibition of the interaction between the ternary complex and TDF that is required for the binding of the ternary complex to 40S subunits.
Partial characterization of a cubic Bacillus phage. Can. J. Microbiol. 24: 986--993. Phage Bam35 is an icosahedron of about 63 nm in diameter. It has a double capsid with spikes at the vertices, and a tail which seems to appear upon nucleic acid ejection. The phage contains DNA and, probably, lipids which seem to be located in the inner coat. The phage is Bacillus-specific, UV- and lipase-resistant, and sensitive to heat, chloroform, and ether. The latent period is 50 min and the burst size is 39. Page Bam35 belongs to a new virus group which includes a phage of B. anthracis and four phages of gram-negative bacteria harboring drug-resistance plasmids.
Six patients with recurrent bladder papillomata were treated both by abdominal scarification with bacillus Calmette-Guérin (BCG) and by intravesical BCG, without resection. Four patients responded favorably (two showing major systemic complications), and the other two, who failed to respond, underwent further surgery.
Explore the source record for details and available documents.
Explore the source record for details and available documents.
Explore the source record for details and available documents.
In ten supine anesthetized dogs, recordings of left atrial (LA) and pulmonary-artery wedge (PW) pressures were simultaneously obtained at several levels of positive end-expiratory pressure (PEEP) ventilation with the thorax either open or closed. Lateral roentgenograms were taken to determine the relative vertical positions of the LA and PW catheter tips. When the wedge catheter tip was vertically above the left artrial catheter tip, mean PW followed airway pressure at PEEP of more that 5 cm H20. For PEEP 5 cm H20 or less, and for PW catheter tip positions vertically below the LA catheter tip at all levels of PEEP, mean wedge catheter pressure was close to left atrial pressure. Thus, it appears the LA pressure can best be estimated by PW catheter positions vertically below the left atrium during positive end-expiratory pressure ventilation.
Explore the source record for details and available documents.
Lymphocytes from the peripheral blood, thoracic duct, spleen and lymph nodes of normal, thymectomized (TX) and thymectomized lethally irradiated marrow reconstituted (TXBM or B rats) Lewis rats were studied for their ability to proliferate in vitro in the presence of Con A or PHA. At the same time the lymphoid tissues of these animals were examined for the presence of B cells or T cells by immunofluorescence staining with antiserums to rat immunoglobulins or rat brain antigens (ARBS), respectively. The specificity of ARBS fort T cells was first established in both cytotoxicity and immunofluoresecnce studies with thymocytes as well as pruified T and B cells. In various lymphoid tissues from TX and B rats, cells carrying brain antigens were found which were unable to respond to Con A and PHA. Thymectomy has a lesser effect on the response to Con A that to PHA; this was particularly true in the thoracic duct where, following thymectomy, lymphocytes continued to respond to Con A but lost their ability to respond to PHA. This suggests that the response to PHA and Con A may belong to different cells' subsets but that the ability to respond to either one of these mitogens may be impaired in cells carrying known T cell markers.
Immunologic approaches to studying lipoprotein structure have been limited because the methods have not been quantitative enough. Recently we reported (Schonfeld, G., and Pfleger, B. (1974) J. Clin. Invest. 54, 236-246) a radioimmunoassay for human apoprotein A-1 (ApoA-I). Only 8% of the ApoA-I of high density lipoprotein (HDL) reacted in the radioimmunoassay system consisting of rabbit anti-human ApoA-I, 125I-ApoA-I, and unlabeled ApoA-I. We suggested that the ApoA-I in HDL were poorly reactive in the radioimmunoassay because they were "masked" by lipid- or protein-protein interactions. To test this, "lipoproteins" were reconstituted from lipids and apoproteins and assayed for their reactivity in the radioimmunoassay. Apo-HDL, ApoA-I alone, or ApoA-I + ApoA-II were sonified with lecithin or with lipids extracted from HDL. Sonicates were fractionated by ultracentrifugation or by Sepharose 4B chromatography. HDLs were also made by incubating dispersed lecithin or lecithin + cholesterol with Apo-HDL, ApoA-I, or ApoA-II. The lipoproteins were analyzed for lipids and protein chemically. Apoprotein compositions were determined by polyacrylamide disc gel electrophoresis. ApoA-I content by radioimmunoassay then was compared with the ApoA-I content obtained by disc gel electrophoresis. Most reconstituted "lipoproteins" had less than the expected ApoA-I contents. Discrepancies between ApoA-I contents were greatest for lipoproteins prepared from Apo-HDL and HDL-lipids (20 to 30% of expected contents). Discrepancies were smaller for particles prepared with lecithin, with ApoA-I alone or with ApoA-I + ApoA-II (40 to 85% of expected). HDLs made by incubation were less reactive than those prepared by sonication. Thus, the reactivity of ApoA-I in the radioimmunoassay could be diminished by causing it to interact with lipids or their apoproteins, or both, suggesting that antigenic sites became masked. From this one can extrapolate that the poor reactivity of the ApoA-I in HDL isolated from plasma also may have been due to the masking of some of its antigenic determinants. The identification of the determinants involved awaits the development of radioimmunoassays for specific regions of ApoA-I.
Morphological and histopathological heart changes were determined for sixteen dystrophic Syrian hamsters (B10 14.6 strain) and sixteen normal hamsters. Eight animals were randomly assigned to each of the following groups: dystrophic swim (DYS-SWM), dystrophic sedentary (DSY-SED), normal swim (NOR-SWM), and normal sedentary (NOR-SED). The daily swimming program consisted of an initial 30-minute swim which was gradually extended to 60 minutes by the end of eight weeks. Weights up to 3% body weight were attached during swimming to increase the work load. Sedentary animals received no experimental treatment. Four animals in each group were sacrificed at 4 and 8 weeks after the initiation of treatments. In comparison with the two groups of sedentary animals, the NOR-SWM group had a greater heart weight/body weight ratio at both 4 and 8 weeks (P less than .05), while the DYS-SWM group had an increased ratio only at 8 weeks (P less than .05). Subjective histopathological evaluation of heart lesions showed that the DYS-SED group had many large areas of inflammatory reaction with infrequent diffuse areas of calcification. In contrast, the DYS-SWM group had fewer and smaller areas of inflammatory reaction with moderate amounts of calcification.
Explore the source record for details and available documents.
Explore the source record for details and available documents.