Calf thymus ribonuclease H IIa activity lacks ribonuclease H specificity.

Less purified fractions of ribonuclease H IIa activity of calf thymus display divalent cation-dependent ribonuclease H activity and divalent cation-independent ribonuclease activity. Because the ratio of the two enzyme activities does not change during successive chromatographic procedures, we suggest that ribonuclease H IIa activity is indeed able to degrade both ssRNA and the RNA moiety of RNA.DNA-hybrids. Ribonuclease H IIa activity can therefore be differentiated from calf thymus ribonuclease H I and H IIb by its lack of ribonuclease H specificity. The native molecular mass of ribonuclease H IIa activity is between 23 and 28 kDa. Under denaturing conditions a 23 kDa-protein band copurifies with the enzyme activity suggesting that this enzyme is monomeric.     

Calf thymus ribonuclease H IIa activity lacks ribonuclease H specificity

Summary Less purified fractions of ribonuclease H IIa activity of calf thymus display divalent cation-dependent ribonuclease H activity and divalent cation-independent ribonuclease activity. Because the ratio of the two enzyme activities does not change during successive chromatographic procedures, we suggest that ribonuclease H IIa activity is indeed able to degrade both ssRNA and the RNA moiety of RNA·DNA-hybrids. Ribonuclease H IIa activity can therefore be differentiated from calf thymus ribonuclease H I and H IIb by its lack of ribonuclease H specificity. The native molecular mass of ribonuclease H IIa activity is between 23 and 28 kDa. Under denaturing conditions a 23 kDa-protein band copurifies with the enzyme activity suggesting that this enzyme is monomeric.     

Detection and localization of a ribonuclease activity associated with vaccinia virus]

A ribonuclease associated with vaccinia virus can be detected when reduced concentrations of nucleotides are used for an in vitro RNA synthesis assay. The non-viral origin of this ribonuclease may be inferred from its external location and from its variable activity on different purified virus stocks. The detection of this ribonuclease activity on purified virus grown without foetal Calf serum may suggest that this enzyme is of cellular origin.     

Class I and class II ribonuclease H activities in Crithidia fasciculata (Protozoa).

The protozoan Crithidia fasciculata contains two different ribonuclease H activities. These enzymes display similar physical and biochemical characteristics to their homologues in higher eukaryotes, for instance calf thymus class I and class II ribonuclease H. Class I ribonuclease H of lower and higher eukaryotes can be activated by Mg2(+)- and Mn2(+)-ions. However, the presence of Mn2(+)-ions is inhibitory for the Mg2(+)-dependent class II ribonuclease H activity of Crithidia fasciculata and calf thymus. The protozoan class I-homologue enzyme appears to be serologically related to the class I ribonuclease H of calf thymus.     

Level of polysomes in cells of Escherichia coli treated with sublethal doses of aminoglycosides

Polysome level was measured in Escherichia coli cells grown in the presence of sublethal doses of four different aminoglycosides which all induce mistranslation: streptomycin, neomycin, kanamycin and gentamycin. No significant variation was noticed as referred to the level of polysomes in normally growing cells. This observation indicates that the erroneous incorporation of amino acids into polypeptide chains does not necessarily result in the release of ribosomes from messenger RNA molecules.     

Effect of exercise on ribonuclease activity in rat skeletal muscle.

Distribution of ribonuclease activity (measured at pH 7.6) in subcellular fractions of homogenates of rat skeletal muscle was investigated in sedentary animals and after 8 weeks running program. Training increased ribonuclease activity (expressed as units of enzyme per g of muscle protein). There was no increase in nuclear fraction, but in both cytoplasmic and mitochondrial fractions the RNA-ase activity increased 42% and 45% respectively.     

Class I and class II ribonuclease H activities inCrithidia fasciculata (protozoa)

Summary The protozoanCrithidia fasciculata contains two different ribonuclease H activities. These enzymes display similar physical and biochemical characteristics to their homologues in higher eukaryotes, for instance calf thymus class I and class II ribonuclease H. Class I ribonuclease H of lower and higher eukaryotes can be activated by Mg²⁺- and Mn²⁺- ions. However, the presence of Mn²⁺-ions is inhibitory for the Mg²⁺-dependent class II ribonuclease H activity ofCrithidia fasciculata and calf thymus. The protozoan class I-homologue enzyme appears to be serologically related to the class I ribonuclease H of calf thymus.     

Adenylate cyclase and phosphodiesterase from brain tissue: different stabilities during incubation of cerebral cortical slices.

Adenylate cyclase and phosphodiesterase were prepared from brain slices from guinea-pig. The specific activity of adenylate cyclase declined rapidly with increasing incubation time of tissue slices, while phosphodiesterase activity was almost uneffected by the incubation of brain slices.     

Activity of two components of serum ribonuclease under conditions of physical exercise.

Serum ribonuclease activity before and after physical exercise in healthy persons was estimated. It is found that a work load of 6000 kgm/5 min increased ribonuclease activity measured at pH 8.5 and decreased the activity of the same enzyme measured at pH 7.0 in the presence of Zn SO4. The observed changes were more pronounced in untrained than in trained persons.     

Adenylate cyclase and phosphodiesterase from brain tissue: Different stabilities during incubation of cerebral cortical slices

Summary Adenylate cyclase and phosphodiesterase were prepared from brain slices from guinea-pig. The specific acitivity of adenylate cyclase declined rapidly with increasing incubation time of tissue slices, while phosphodiesterase activity was almost uneffected by the incubation of brain slices.     

Effect of exercise on ribonuclease activity in rat skeletal muscle

Summary Distribution of ribonuclease activity (measured at pH 7.6) in subcellular fractions of homogenates of rat skeletal muscle was investigated in sedentary animals and after 8 weeks running program. Training increased ribonuclease activity (expressed as units of enzyme per g of muscle protein). There was no increase in nuclear fraction, but in both cytoplasmic and mitochondrial fractions the RNA-ase activity increased 42% and 45% respectively.Acknowledgments. I would like to thank Prof. L. elewski and Dr J. Popinigis for helpful suggestions.     

Activity of two components of serum ribonuclease under conditions of physical exercise

Summary Serum ribonuclease activity before and after physical exercise in healthy persons was estimated. It is found that a work load of 6000 kgm/5 min increased ribonuclease activity measured at pH 8.5 and decreased the activity of the same enzyme measured at pH 7.0 in the presence of ZnSO₄. The observed changes were more pronounced in untrained than in trained persons.     

The cytotoxicity of eosinophil cationic protein/ribonuclease 3 on eukaryotic cell lines takes place through its aggregation on the cell membrane

Human eosinophil cationic protein (ECP)/ ribonuclease 3 (RNase 3) is a protein secreted from the secondary granules of activated eosinophils. Specific properties of ECP contribute to its cytotoxic activities associated with defense mechanisms. In this work the ECP cytotoxic activity on eukaryotic cell lines is analyzed. The ECP effects begin with its binding and aggregation to the cell surface, altering the cell membrane permeability and modifying the cell ionic equilibrium. No internalization of the protein is observed. These signals induce cell-specific morphological and biochemical changes such as chromatin condensation, reversion of membrane asymmetry, reactive oxygen species production and activation of caspase-3-like activity and, eventually, cell death. However, the ribonuclease activity component of ECP is not involved in this process as no RNA degradation is observed. In summary, the cytotoxic effect of ECP is attained through a mechanism different from that of other cytotoxic RNases and may be related with the ECP accumulation associated with the inflammatory processes, in which eosinophils are present. Received 26 October 2007; accepted 23 November 2007     

Degradation of ribosomal RNA from Escherichia coli by ribonuclease U2 and in vitro transcription of "RNA-fragments" into complementary DNA]

Under well defined conditions ribosomal RNAs purified from Escherichia coli can be degraded by ribonuclease U2 giving rise to RNA fragments of 60--70 nucleotides. In vitro, these fragments are efficiently transcribed into a complementary DNA by DNA polymerase RNA dependent, partially purified from extracts of E. coli. In vivo, "RNA-fragments-U2" inhibit the development of plant tumors.     

Opposite effect of PGE2 on cAMP levels in human adrenal medulla and pheochromocytoma.

PGE2 (10(-7) M) caused increased cAMP accumlation in 5 pheochromocytomas, while in 3 human adrenal medullae PGE2 caused a significant decrease of cAMP level on incubating slices in vitro. This finding is discussed in relation to the opposite effect of PGE2 on catecholamine release from human medulla and pheochromocytoma slices in vitro.     

The eosinophil ribonucleases

The eosinophil ribonucleases, eosinophil-derived neurotoxin (EDN/RNase 2) and eosinophil cationic protein (ECP/RNase 3) are two closely related proteins with intriguing functional and evolutionary properties. While both EDN and ECP maintain the structural and catalytic residues typical of the RNase A superfamily, the role of ribonuclease activity in the physiologic function of these proteins remains unclear. The biochemistry and physiology of EDN, ECP and the recently discovered ribonuclease k6 (RNase 6) will be reviewed in this chapter.     

Differential alterations in branched-chain amino acid decarboxylation in liver of hypophysectomized rats.

Valine decarboxylation was significantly increased and leucine decarboxylation was significantly decreased in rat liver slices following hypophysectomy. In both normal and hypophysectomized rats decarboxylation of leucine exceeded that of valine in slices whereas the reverse was observed with the respective keto acids and mitochondria.     

Détection cytochimique de l'acide ribonucléique dans les chromosphères pré- et post-méiotiques des sporanges de résistance d'Allomyces

Summary Premeiotic chromospheres in the immature resistant sporangia (RS) ofAllomyces lose their basophily after treatment with trichloracetic acid, perchloric acid or ribonuclease. Postmeiotic chromospheres in the mature RS, becoming the nuclear caps of the ‘reduced’ zoospores, do not stain with toluidine blue or pyronine after ribonuclease digestion. It is concluded that, as gametic and zoosporic nuclear caps, pre- and postmeiotic chromospheres are transitory cytoplasmic organites rich in RNA.      

Limitation de l'emploi des cóupes tissulaires pour l'étude des phénomènes métaboliques

Summary The use of tissues slices for metabolic studies gives results valuable only from the qualitative view-point. The histochemistry of liver slices reveals indeed that there is a disappearance of the basophilia of the inner layers of cells and, consequently, autoradiographic preparations of slices incubated with tagged glycine show that the incorporation of the amino-acid is limited to the outer layers of cells.