Subject(s)
Placental Hormones/pharmacology , RNA, Ribosomal/biosynthesis , Ribonucleases/antagonists & inhibitors , Animals , Carcinoma, Ehrlich Tumor/enzymology , Cell Nucleolus/enzymology , Cell-Free System , Ethylmaleimide/pharmacology , Female , Humans , Kinetics , Mice , Molecular Weight , Pregnancy , RNA, Ribosomal/genetics , Transcription, GeneticABSTRACT
A nucleolar ribonuclease specific for single-stranded ribonucleic acid (RNA) has been isolated and extensively purified from Ehrlich ascites carcinoma cells. The enzyme is optimally active at neutral pH and degrades RNA via a 2',3'-cyclic intermediate leaving 3'- or 2',3'-cyclic terminated oligonucleotides. The ribonuclease has an apparent molecular weight of 38 500 as judged by sedimentation equilibrium and is a basic protein having an isoelectric point greater than 9.0. The enzyme preferentially cleaves poly(C) over poly (U), poly(A), or poly(C).poly(I). Limit digestion products of poly(C) degratation are on the average tri-, tetra-, and pentanucleotides. In the partial digestion of yeast 5.8S rRNA, the nucleolar ribonuclease cleaves only CpA phosphodiester bonds. Spermidine, spermine, and histone I inhibit the activity of nucleolar ribonuclease. Antibodies directed toward pancreatic RNase do not cross-react with the Ehrlich nucleolar ribonuclease.
Subject(s)
Carcinoma, Ehrlich Tumor/enzymology , Cell Nucleolus/enzymology , Ribonucleases/metabolism , Animals , Base Sequence , Immunoassay , Kinetics , Mice , Polyribonucleotides , Ribonucleases/isolation & purification , Substrate SpecificityABSTRACT
The rate of uptake of uridine into the acid-soluble fraction of Novikoff hepatoma cells is inhibited by low concentrations of the ionophores A23187 and gramicidin and other perturbants of intracellular cation levels. Inhibition of uridine uptake by A23187 is dependent on Ca2+ and is reduced by serum and high levels of Mg2+. The effectiveness of A23187 is dependent on the Ca2+/Mg2+ ratio rather than the absolute concentration of either ion. Inhibition of uridine uptake by gramicidin is not significantly affected by serum or divalent cations. Other effectors of monovalent cation flux such as ouabain and valinomycin also inhibit uridine uptake. These results indicate that net uptake of uridine may be influenced by intracellular levels of certain monovalent and divalent inorganic cations.
