A truncation in the 14 kDa protein of the signal recognition particle leads to tertiary structure changes in the RNA and abolishes the elongation arrest activity of the particle.
The signal recognition particle (SRP) provides the molecular link between synthesis of polypeptides and their concomitant translocation into the endoplasmic reticulum. During targeting, SRP arrests or delays elongation of the nascent chain, thereby presumably ensuring a high translocation efficiency. Components of the Alu domain, SRP9/14 and the Alu sequences of SRP RNA, have been suggested to play a role in the elongation arrest function of SRP. We generated a truncated SRP14 protein, SRP14-20C, which forms, together with SRP9, a stable complex with SRP RNA. However, particles reconstituted with SRP9/14-20C, RC(9/14-20C), completely lack elongation arrest activity. RC(9/14-20C) particles have intact signal recognition, targeting and ribosome binding activities. SRP9/14-20C therefore only impairs interactions with the ribosome that are required to effect elongation arrest. This result provides evidence that direct interactions between the Alu domain components and the ribosome are required for this function. Furthermore, SRP9/14-20C binding to SRP RNA results in tertiary structure changes in the RNA. Our results strongly indicate that these changes account for the negative effect of SRP14 truncation on elongation arrest, thus revealing a critical role of the RNA in this function.
Thomas
Y
Y
Département de Biologie Cellulaire, Université de Genève, Sciences III, CH-1211 Genève 4, Switzerland.
Bui
N
N
Strub
K
K
D000595:Amino Acid Sequence; D000818:Animals; D003001:Cloning, Molecular; D016384:Consensus Sequence; D019281:Dimerization; D004285:Dogs; D004926:Escherichia coli; D017665:Hydroxyl Radical; D007700:Kinetics; D051379:Mice; D008969:Molecular Sequence Data; D009690:Nucleic Acid Conformation; D010944:Plants / Q000378:metabolism; D016133:Polymerase Chain Reaction; D012335:RNA, Ribosomal; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000378:metabolism; D012091:Repetitive Sequences, Nucleic Acid; D012441:Saccharomyces cerevisiae / Q000378:metabolism; D017384:Sequence Deletion; D018271:Signal Recognition Particle / Q000096:biosynthesis / Q000737:chemistry / Q000378:metabolism*
D012335:RNA, Ribosomal; D011994:Recombinant Proteins; D018271:Signal Recognition Particle; C503637:Srp72 protein, mouse; D017665:Hydroxyl Radical