A phylogenetic analysis of Legionella

Four species of Legionella, L. pneumophila NCTC 11192, L. bozemanii NCTC 11368, L. micdadei NCTC 11371 and L. jordanis ATCC 33623 have been characterized by oligonucleotide cataloguing of their 16S ribosomal RNA. All four species are phylogenetically closely related, while no specific relationship could be detected with any other group of organisms investigated so far with respect to this method. At a low level of relationship legionellae are members of the broad group of purple photosynthetic bacteria and their non-phototrophic relatives, in which Legionella form an independent line of descent.     

Proteins of the sieve-tube exudate of Cucurbita maxima

Summary Sieve-tube exudate which appears on cut surfaces of stems of Cucurbita maxima as distinct droplets has been depicted in electron micrographs of longitudinal sections of the phloem. The exudate, which was produced from mature sieve tubes only, contained filaments of P-protein, but no mitochondria or vesicles of endoplasmic reticulum. The water-soluble part of the exudate contained at least 12 proteins, as shown by disc-electrophoresis. Enzymic activity was found for peroxidases, acid phosphatases, and aldolases. Color tests and assays for other enzymes, including ATPase, fructokinase, several dehydrogenases, and UDP-glucose: D-fructose-2-glucosyl transferase, gave negative results. With repeated cutting of a stem, the protein content of the exudate increased, while the amount of exudate decreased.Supported by Deutsche Forschungsgemeinschaft and Stiftung Volkswagenwerk. During part of this investigation the senior author held a U.S. National Science Foundation Senior Foreign Scientist Fellowship at the University of Wisconsin.     

Two Pathways of Glutamate Fermentation by Anaerobic Bacteria

Two pathways are involved in the fermentation of glutamate to acetate, butyrate, carbon dioxide, and ammonia-the methylaspartate and the hydroxyglutarate pathways which are used by Clostridium tetanomorphum and Peptococcus aerogenes, respectively. Although these pathways give rise to the same products, they are easily distinguished by different labeling patterns of the butyrate when [4-(14)C]glutamate is used as substrate. Schmidt degradation of the radioactive butyrate from C. tetanomorphum yielded equally labeled propionate and carbon dioxide, whereas nearly all the radioactivity of the butyrate from P. aerogenes was recovered in the corresponding propionate. This procedure was used as a test for the pathway of glutamate fermentation by 15 strains (9 species) of anaerobic bacteria. The labeling patterns of the butyrate indicate that glutamate is fermented via the methylaspartate pathway by C. tetani, C. cochlearium, and C. saccarobutyricum, and via the hydroxyglutarate pathway by Acidaminococcus fermentans, C. microsporum, Fusobacterium nucleatum, and F. fusiformis. Enzymes specific for each pathway were assayed in crude extracts of the above organisms. 3-Methylaspartase was found only in clostridia which use the methylaspartate pathway, including Clostridium SB4 and C. sticklandii, which probably degrade glutamate to acetate and carbon dioxide by using a second amino acid as hydrogen acceptor. High levels of 2-hydroxyglutarate dehydrogenase were found exclusively in organisms that use the hydroxyglutarate pathway. The data indicate that only two pathways are involved in the fermentation of glutamate by the bacteria analyzed. The methylaspartate pathway appears to be used only by species of Clostridium, whereas the hydroxyglutarate pathway is used by representatives of several genera.     

Mechanism of α Factor Biosynthesis in Saccharomyces cerevisiae

The biosynthesis of alpha factor, a mating-type-specific regulatory oligopeptide which is secreted by Saccharomyces cerevisiae cells of alpha mating type, was studied. In batch cultures only small amounts of the peptide were synthesized during the exponential growth phase. During the stationary phase, alpha factor was produced at a constant rate and accumulated in the culture medium. Inhibition of translation in wild-type cells by cycloheximide, or in mutant strains under conditions which blocked protein or ribonucleic acid (RNA) synthesis completely inhibited the production of alpha factor. These results indicate that the factor is produced by ribosomal translation of a specific messenger RNA and not by an extraribosomal mechanism of peptide synthesis.     

The goldfish pituitary

Summary In the goldfish, Carassius auratus, morphological and functional aspects of the pituitary gland were studied at the ultrastructural level and six cell types could be distinguished in the pars distalis. Acidophilic cells of the rostral pars distalis were identified as prolactin cells, the chromophobic cells of the rostral pars distalis as ACTH cells, the non-globular basophilic cells of the rostral and the proximal pars distalis as TSH cells, the globular basophils of the proximal pars distalis as gonadotropic cells and the acidophils of the proximal pars distalis as somatotrophs.Besides some of the well established criteria of morphological and functional identification of different cell types, two new approaches have been used in the present study. One was to express the electron density of secretory granules objectively by means of a photometric method. It was found that both types of acidophilic cells which produce the proteohormones prolactin and somatotropin respectively, had granules with the highest electron densities. The basophilic cells producing the glycoproteins gonadotropin and TSH respectively, possessed granules of intermediate electron density whereas the chromophobic cells storing the peptide hormone ACTH had granules of lowest densities. The second new approach was the administration of the synthetic mammalian releasing hormones LH-RP and TRF, which helped in identifying gonadotropic and thyrotropic cells respectively. In the goldfish there is evidence for the presence of only one type of gonadotropic cell.Supported by a grant of the Science Research Council of Great Britain to Professor Sir Francis Knowles, F.R.S. The electron microscope used was provided by the Medical Research Council of Great Britain. The integrating photometer IPM2 was kindly on loan from Messrs. Carl Zeiss, Oberkochen, Germany. For technical advice we are greatly indebted to Mr. P. K. Kaul, B. E., M.I. Struct. E., C. Eng.     

Stimulation by 6-N,2′-O-dibutyryladenosine 3′:5′-cyclic monophosphate and glucocorticoids of phosphoenolpyruvate carboxykinase in the isolated perfused rat liver (Short Communication)

Dibutyryl cyclic AMP stimulated the activity of phosphoenolpyruvate carboxykinase in perfused livers of rats, fed on a low-protein diet, linearly over a 6h period. The enzyme activity was also significantly elevated by dexamethasone, the effect being considerably lower than that of the cyclic nucleotide. Since the time-course of phosphoenolpyruvate carboxykinase activity in response to dibutyryl cyclic AMP resembled that observed after dibutyryl cyclic AMP injection into intact animals, it is suggested that induction of the enzyme in vivo is due to a direct action of the cyclic nucleotide on the liver. Combined administration of dibutyryl cyclic AMP and glucocorticoids did not lead to an additive increase of liver phosphoenolpyruvate carboxykinase activity, either in vivo or in the perfused organ.