Heterogeneous nuclear ribonucleoprotein A1 catalyzes RNA.RNA annealing.
Within the nucleus, pre-mRNA molecules are complexed with a set of proteins to form heterogeneous nuclear ribonucleoprotein complexes. A1, an abundant RNA binding protein present in these complexes, has been shown to bind selectively to single-stranded RNAs and destabilize base-pairing interactions. In this study.A1 is shown to promote the rate of annealing of complementary RNA strands greater than 300-fold under a wide range of salt concentration and temperature. Maximal annealing is observed under saturating or near saturating concentrations of protein, but annealing decreases sharply at both higher and lower concentrations of A1. Kinetic analysis shows that the rate of annealing is not strictly first or second order with respect to RNA at a ratio of protein/RNA that gives optimal rates of annealing. This result suggests that A1 protein may affect more than one step in the annealing reaction. Two polypeptides representing different domains of A1 were also examined for annealing activity. UP1, a proteolytic fragment that represents the N-terminal two-thirds of A1, displays very limited annealing activity. In contrast, a peptide consisting of 48 amino acid residues from the glycine-rich C-terminal region promotes annealing at a rate almost one-quarter that observed with intact A1. The RNA.RNA annealing activity of A1 may play a role in pre-mRNA splicing and other aspects of nuclear mRNA metabolism.
Department of Biology, Marquette University, Milwaukee, WI 53233.
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D000076245:Heterogeneous Nuclear Ribonucleoprotein A1; D034481:Heterogeneous-Nuclear Ribonucleoprotein Group A-B; D034441:Heterogeneous-Nuclear Ribonucleoproteins; D007700:Kinetics; D015636:Magnesium Chloride / Q000737:chemistry; D009693:Nucleic Acid Hybridization; D010455:Peptides / Q000737:chemistry; D012313:RNA / Q000737:chemistry*; D012261:Ribonucleoproteins / Q000502:physiology*; D012965:Sodium Chloride / Q000737:chemistry; D013329:Structure-Activity Relationship; D013696:Temperature
D000076245:Heterogeneous Nuclear Ribonucleoprotein A1; D034481:Heterogeneous-Nuclear Ribonucleoprotein Group A-B; D034441:Heterogeneous-Nuclear Ribonucleoproteins; D010455:Peptides; D012261:Ribonucleoproteins; D015636:Magnesium Chloride; D012965:Sodium Chloride; D012313:RNA