Immunological study of the regulation of cellular arylsulfatase synthesis in Klebsiella aerogenes.

Regulation of cellular arylsulfatase synthesis in Klebsiella aerogenes was analyzed by immunological techniques. Antibody directed against the purified arylsulfatase from K. aerogenes W70 was obtained from rabbits and characterized by immunoelectrophoresis, double-diffusion, quantitative precipitation, and enzyme neutralization tests. Arylsulfatase was located in the periplasmic space when the wild-type strain was cultured with methionine or with inorganic sulfate plus tyramine, but not with inorganic sulfate without tyramine, as the sole sulfur source. Tyramine oxidase was retained in the membrane fraction prepared from cells grown in the presence of tyramine. Arylsulfatase protein was not synthesized in the presence of tyramine and inorganic sulfate by mutant K611, which is deficient in tyramine oxidase (tynA). We conclude that the expression of the arylsulfatase gene (atsA) is regulated by the expression of tynA and that inorganic sulfate serves as a corepressor. In addition, strains mutated in the atsA gene were analyzed by using antibody.     

Formation of regular packets of Staphylococcus aureus cells.

Staphylococcus aureus, which usually forms grape-like clusters, has the ability to form regularly arranged cell packets. These regular cell packets are formed when the activity of its separation enzyme(s) is lost either by treatment with detergents, such as sodium dodecyl sulfate or Trition X-100, or by mutation of the cells. These cell packets consisted of 8 to 64 spherical cells that have a three-dimensional arrangement. Some irregularity in the arragement of cells in packets, however, can be observed by scanning electron microscopy. It is concluded that S. aureus fundametally divides along three definitely oriented planes that are located at right angles to each other. After cell division, the cells usually become translocated due to the action of a separation enzyme(s) to form grape-like clusters.     

Oxidation of linear terpenes and squalene variants by Arthrobacter sp.

Cells of Arthrobacter sp. that had been isolated from soil were used to study oxidation of some linear terpenes and squalene variants. The cells oxidized geraniol, nerol, and farnesol to the corresponding aldehydes, with partial conversion of the geometrical isomerism of the alpha,beta-double bond. The squalene variant, squalene-2,3-oxide, was cleaved to 9,10-epoxygeranylacetone and geranylacetone. Squalene-2,3-22,23-dioxide was cleaved to 9,10-epoxygeranylacetone. These products were optically active, and their stereochemistry and optical purity were determined.     

The active form of phytochrome: a new hypothesis based on phytochrome pelletability studies

Difficulties arising from the current dogma that the far-red absorbing form of phytochrome (P_fr) is the only active form are discussed.A new hypothesis is proposed in which phytochrome is held to be the photoreceptor for both low energy (pulse) and high energy (HIR) responses. There is a common basic mechanism of action involving interaction between phytochrome and a binding site within the cell. The phytochrome involvement in low energy responses exhibits an action spectrum for binding that matches the P_r absorption spectrum and reversibility by far-red irradiation. Upon prolonged irradiation the phytochrome-binding site interaction acquires different characteristics that are reminiscent of those displayed in HIR, e.g. dependence on sustained irradiation for continual binding, dependence of the degree of binding on irradiance and the similarity of the action spectrum with that of HIR action spectra, e.g. that for inhibition of lettuce hypocotyl lengthening.As expected on the basis of the new hypothesis the particulate fraction of phytochrome contains both P_r and P_fr. Arguments are advanced that the presence of P_r in pellets of particulate phytochrome cannot be accounted for by (i) the “induced fit” hypothesis, (ii) the “pigment cycling” hypothesis, and (iii) the “open phytochrome-receptor model”. We conclude that phytochrome molecules, after being sufficiently energized can interact with their intracellular binding sites irrespective of their chromophoric configuration.     

Presence of somatostatin-28 like immunoreactivity in the human pancreas

Specific antisera against somatostatin-28 were prepared by absorption of somatostatin-28 antisera with sepharose 4B-somatostatin-14. Indirect immunofluorescence techniques using somatostatin-14 antisera and specific antisera against somatostatin-28 were carried out to elucidate the time of occurrence of somatostatin-28 in the fetal pancreatic islets and to ascertain whether somatostatin-28 was present in the adult pancreatic islets or not, and further to examine whether cells reacting with specific antisera against somatostatin-28 are identical to those reacting with somatostatin-14 antisera or not. Somatostatin-28 like immunoreactivity occurred in the fetal pancreatic islets at 11th week's gestation and was found in all fetal pancreatic islets examined in the present study. It was also found in the adult pancreatic islets. Furthermore, cells reacting with specific antisera against somatostatin-28 in the fetal and adult pancreatic islets were identical to those reacting with somatostatin-14 antisera. Thus, the present study elucidated the presence of somatostatin-28 like immunoreactivity in the human pancreas. However, it could not be decided whether cells reacting with somatostatin-28 antisera contain either only somatostatin-28 or both somatostatin-28 and somatostatin-14; in other words, whether somatostatin-14 is produced from somatostatin-28 or not, since somatostatin-14 antisera had a cross-reactivity to both somatostatin-14 and somatostatin-28.     

Molecular cloning of the ribosomal RNA genes of the photosynthetic bacterium Rhodopseudomonas capsulata

Summary Chromosomal segments of Rhodopseudomonas capsulata carrying the ribosomal operons and cloned with the cosmid vector pHC79 have been identified by cross hybridization with ³²P-ATP labeled rRNAs. At least seven rRNA operons are present in the R. capsulata chromosome. By R-loop analyses of DNA-RNA hybrids, two distinct loop structures of sizes 1.50 kb and 2.52 kb corresponding to the 16S and 23S RNA molecules, respectively, were detected. Intact 23S RNA molecules can be isolated from R. capsulata ribosomes by sucrose density centrifugation. However, fragmentation of the 23S RNA molecule into a 16S-like molecule was observed during gel electrophoresis. Restriction mapping and hybridization of a 9 kb PstI fragment that contained one copy of the rRNA operon showed the following sequence of the RNA genes in R. capsulata 16S, 23S, and 5S. A spacer region of 0.91 kb was found between the 16S and the 23S RNA genes.     

Analysis of long-latency components of field potentials evoked by stimulation of the cerebral cortex and nerves in the cerebellar paramedian lobule

Field potentials evoked in the graunular layer of the cerebellar paramedian lobule of unanesthetized cats in response to stimulation of the sensomotor cortex and limb nerves contained slow negative waves, appearing after a long latent period, which were generated by granule cells. In the case of nerve stimulation this component was recorded both inside and outside the projection zone of the corresponding limb. Cortical stimulation by single stimuli or series of stimuli not more than 1.8–2.5 times above threshold strength led to the appearance of evoked potentials only inside the corresponding projection zone. The long-latency component of field potentials evoked by cerebral stimulation followed high frequencies of repetitive stimulation and was less sensitive to the action of barbital anesthesia than the analogous component of potentials evoked by nerve stimulation. In the case of combined cerebral and nerve stimulation the long-latency components underwent summation. It is concluded that mossy fibers of slowly-conducting spino- and cerebrocerebellar tracts innervate different granule cells in the cerebellar cortex.Institute of Problems in Information Transmission, Academy of Sciences of the USSR, Moscow. M. V. Lomonosov Moscow State University. Translated from Neirofiziologiya, Vol. 14, No. 4, pp. 379–385, July–August, 1982.     

Ion exchange in the horizontal cells of the retina

Experimental data indicate that the membrane potential of L-type horizontal cells of the retina to bright light (i.e., when synaptic inputs are completely closed) is close to the potassium equilibrium potential. From this observation the intracellular concentration of K⁺ and Na⁺ was estimated. The latter was found to be relatively high (tens of millimoles/liter), i.e., comparable with the intracellular K⁺ concentration. This result, coupled with data on closeness of the equilibrium potential of the photic response to zero, is evidence that besides sodium conductance, the potassium conductance of the subsynaptic membrane also participates in generation of the photic response by these cells. The steady-state sodium and potassium synaptic currents was shown to be relatively small and to vary only a little over the whole working range of potentials (from –72 to –16 mV), due to the nonlinear properties of the nonsynaptic cell membrane.Institute for Problems in Information Transmission, Academy of Sciences of the USSR, Moscow. Translated from Neirofiziologiya, Vol. 14, No. 1, pp. 3–10, January–February, 1982.     

The selection of cultured plant cell lines producing high levels of biotin

We have found that biotin is synthesized in many species of cultured plant cells, e.g. Lavandula vera Labiatae), Nicotiana tabacum (Solanaceae) and Glycine max Leguminosae). Cultured green L. vera cells grown under light contained the greatest amounts of free biotin of the cells studied although the specific amounts varied among the cell lines. Cell lines were selected after their free biotin contents had been analysed. Cells containing large amounts of free biotin were cultured repeatedly, analysed and reselected. Lines with high levels of free biotin were obtained from cells which survived on a medium containing pimelic acid and l-alanine or from gamma irradiated cells. One L. vera cell line obtained from irradiated cells contained seven times the amount of free biotin found in the original unselected cultured cells and four and a half times that found in the leaves.     

An improved method for the isolation of adipocyte plasma membranes.

An improved procedure is outlined for the isolation of an adipocyte plasma membrane fraction containing much less endoplasmic reticulum contamination than plasma membranes prepared by the procedures that are currently in commen use. It is also shown that ¹²⁵I-labeled diazotized diiodosulfanilic acid can be used as a nonpermeable reagent which selectively labels two protein components in plasma membranes of intact adipocytes.