Hierarchical assembly of the Alu domain of the mammalian signal recognition particle.
The mammalian signal recognition particle (SRP) catalytically promotes cotranslational translocation of signal sequence containing proteins across the endoplasmic reticulum membrane. While the S-domain of SRP binds the N-terminal signal sequence on the nascent polypeptide, the Alu domain of SRP temporarily interferes with the ribosomal elongation cycle until the translocation pore in the membrane is correctly engaged. Here we present biochemical and biophysical evidence for a hierarchical assembly pathway of the SRP Alu domain. The proteins SRP9 and SRP14 first heterodimerize and then initially bind to the Alu RNA 5' domain. This creates the binding site for the Alu RNA 3' domain. Alu RNA then undergoes a large conformational change with the flexibly linked 3' domain folding back by 180 degrees onto the 5' domain complex to form the final compact Alu ribonucleoprotein particle (Alu RNP). We discuss the possible mechanistic consequences of the likely reversibility of this final step with reference to translational regulation by the SRP Alu domain and with reference to the structurally similar Alu RNP retroposition intermediates derived from Alu elements in genomic DNA.
Weichenrieder
O
O
European Molecular Biology Laboratory, Grenoble Outstation, France.
Stehlin
C
C
Kapp
U
U
Birse
D E
DE
Timmins
P A
PA
Strub
K
K
Cusack
S
S
D020087:Alu Elements; D019943:Amino Acid Substitution; D000818:Animals; D001483:Base Sequence; D008322:Mammals; D008958:Models, Molecular; D008969:Molecular Sequence Data; D016296:Mutagenesis; D009690:Nucleic Acid Conformation; D012313:RNA / Q000737:chemistry* / Q000378:metabolism; D016601:RNA-Binding Proteins / Q000737:chemistry* / Q000378:metabolism; D012261:Ribonucleoproteins / Q000378:metabolism; D018271:Signal Recognition Particle / Q000737:chemistry* / Q000378:metabolism
D016601:RNA-Binding Proteins; D012261:Ribonucleoproteins; D018271:Signal Recognition Particle; C490325:Srp14 protein, mouse; C503637:Srp72 protein, mouse; C107410:Srp9 protein, mouse; D012313:RNA