Activation of ribonuclease L by (2'-5')(A)4-poly(L-lysine) conjugates in intact cells.
Molecular hybrids were synthesized by coupling (2'-5')(A)n oligoadenylates or 2-5A, an intracellular mediator involved in antiviral activity of interferons (IFNs), with poly(L-lysine) used as a membrane carrier. (2'-5')(A)n in its free form was not taken up by cells, probably because of its ionic character. Conjugation with the polypeptide carrier overcame this problem and enabled its pharmacological properties to be developed. The alpha-glycol group of individual (2'-5')(A)n oligomers was oxidized by periodate oxidation and conjugated by an amino reductive reaction to poly(L-lysine), Mr 14 000, in a molar ratio of 5:1. These hybrid molecules left the biologically active 5' end moiety of the (2'-5')(A)n molecule unchanged, and in particular its triphosphate group, and stabilized the molecule by increasing its resistance to phosphodiesterase hydrolysis. A dose-dependent inhibition of virus growth was observed on concomitant incubation of (2'-5')(A)n-poly(L-lysine) conjugates with vesicular stomatitis virus infected L1210 cell cultures. This was a result of the activation of the (2'-5')(A)n-dependent endoribonuclease (RNase L) by intracellularly delivered (2'-5')(A)n as in some IFN-treated virus-infected cells. Indeed, (2'-5')(A)n-poly(L-lysine) conjugates bind RNase L effectively as can be seen from their ability to compete with authentic (2'-5')(A)n in a cell-free radiobinding assay. Moreover, (2'-5')(A)n-poly(L-lysine) conjugates promote transient inhibition of protein synthesis and a characteristic cleavage pattern of ribosomal RNAs in intact cells.(ABSTRACT TRUNCATED AT 250 WORDS)
D000227:Adenine Nucleotides / Q000138:chemical synthesis / Q000494:pharmacology*; D000818:Animals; D000998:Antiviral Agents / Q000494:pharmacology; D002472:Cell Transformation, Viral / Q000187:drug effects; D004722:Endoribonucleases / Q000378:metabolism*; D004789:Enzyme Activation; D007202:Indicators and Reagents; D007700:Kinetics; D007739:L Cells / Q000201:enzymology; D051379:Mice; D008970:Molecular Weight; D009843:Oligoribonucleotides / Q000494:pharmacology; D011107:Polylysine / Q000138:chemical synthesis / Q000494:pharmacology* / Q000031:analogs & derivatives; D013329:Structure-Activity Relationship; D014721:Vesicular stomatitis Indiana virus / Q000187:drug effects / Q000235:genetics
D000227:Adenine Nucleotides; D000998:Antiviral Agents; D007202:Indicators and Reagents; D009843:Oligoribonucleotides; C049615:adenylyl-(2'-5')-adenylyl-(2'-5')-adenylyl-(2'-5')-adenylyl-polylysine; D011107:Polylysine; C023505:2',5'-oligoadenylate; D004722:Endoribonucleases; C031956:2-5A-dependent ribonuclease