The major messenger ribonucleoprotein particle protein p50 (YB-1) promotes nucleic acid strand annealing.
p50, a member of the Y-box binding transcription factor family, is tightly associated with eukaryotic mRNAs and is responsible for general translational regulation. Here we show that p50, in addition to its previously described ability to melt mRNA secondary structure, is capable of promoting rapid annealing of complementary nucleic acid strands. p50 accelerates annealing of RNA and DNA duplexes up to 1500-fold within a wide range of salt concentrations and temperatures. Phosphorylation of p50 selectively inhibits DNA annealing. Moreover, p50 catalyzes strand exchange between double-stranded and single-stranded RNAs yielding a product bearing a more extended double-stranded structure. Strikingly, p50 displays both RNA-melting and -annealing activities in a dose-dependent manner; a relatively low amount of p50 promotes formation of RNA duplexes, whereas an excess of p50 causes unwinding of double-stranded forms. Our results suggest that the alteration of nucleic acid conformation is a basic mechanism of the p50-dependent regulation of gene expression.
Skabkin
M A
MA
Institute of Protein Research, Russian Academy of Sciences, Pushchino, Moscow Region 142290, Russia.
Evdokimova
V
V
Thomas
A A
AA
Ovchinnikov
L P
LP
D000818:Animals; D047390:Casein Kinase II; D004247:DNA / Q000502:physiology*; D018076:DNA, Complementary / Q000378:metabolism; D004305:Dose-Response Relationship, Drug; D008957:Models, Genetic; D009696:Nucleic Acids / Q000378:metabolism*; D010766:Phosphorylation; D011489:Protein Denaturation; D017346:Protein Serine-Threonine Kinases / Q000378:metabolism; D017433:Protein Structure, Secondary; D012313:RNA / Q000502:physiology*; D012330:RNA, Double-Stranded / Q000378:metabolism; D016601:RNA-Binding Proteins / Q000378:metabolism* / Q000502:physiology*; D011817:Rabbits; D011994:Recombinant Proteins / Q000378:metabolism; D012097:Repressor Proteins / Q000378:metabolism* / Q000502:physiology*; D012492:Salts / Q000494:pharmacology; D013696:Temperature
D018076:DNA, Complementary; D009696:Nucleic Acids; D012330:RNA, Double-Stranded; D016601:RNA-Binding Proteins; D011994:Recombinant Proteins; D012097:Repressor Proteins; D012492:Salts; C118824:p50 Y-box-binding protein, mammalian; D012313:RNA; D004247:DNA; D047390:Casein Kinase II; D017346:Protein Serine-Threonine Kinases