Mechanistic studies of ribonucleic acid renaturation by a helix-destabilizing protein.
The ability of a nucleic acid helix-destabilizing protein from calf thymus, UP1, to facilitate renaturation of yeast tRNALeu3 and Escherichia coli 5S RNA is shown to be a consequence of the protein's ability to bind stoichiometrically to single-stranded polynucleotide regions. A comparison of the inhibitory effect of different homopolymers on UP1-induced renaturation of tRNALeu3 does not indicate significant base specificity in UP1 binding, and a 3'-5' ribose phosphate polymer devoid of heterocyclic bases inhibits as well as the homopolynucleotides. These inhibition studies also show that UP1 requires polynucleotide segments of at least three phosphate residues to bind. Mg2+ (which is required for the stabilization of native tRNALeu3) dissociates complexes of UP1 with inactive tRNA, and since the RNAs in those complexes lack a substantial amount of secondary structure, it can upon dissociation readily refold into the native structure. A semiquantitative treatment of UP1-RNA interaction is developed that suggests that only a small number (approximately six) of protein molecules are bound to tRNALeu3 in the complex while analysis of the inhibition studies suggests that these UP1 molecules are not bound in a highly cooperative manner.
D000818:Animals; D002417:Cattle; D004265:DNA Helicases / Q000494:pharmacology*; D000076245:Heterogeneous Nuclear Ribonucleoprotein A1; D034481:Heterogeneous-Nuclear Ribonucleoprotein Group A-B; D066298:In Vitro Techniques; D007700:Kinetics; D008274:Magnesium / Q000494:pharmacology; D009695:Nucleic Acid Renaturation / Q000187:drug effects*; D009841:Oligonucleotides / Q000494:pharmacology; D011119:Polynucleotides / Q000494:pharmacology; D012313:RNA; D012329:RNA, Bacterial; D012331:RNA, Fungal; D012346:RNA, Transfer, Amino Acyl; D012261:Ribonucleoproteins; D013951:Thymus Hormones / Q000494:pharmacology*
D000076245:Heterogeneous Nuclear Ribonucleoprotein A1; D034481:Heterogeneous-Nuclear Ribonucleoprotein Group A-B; D009841:Oligonucleotides; D011119:Polynucleotides; D012329:RNA, Bacterial; D012331:RNA, Fungal; D012346:RNA, Transfer, Amino Acyl; D012261:Ribonucleoproteins; D013951:Thymus Hormones; D012313:RNA; D004265:DNA Helicases; D008274:Magnesium