Signal recognition particle Alu domain occupies a defined site at the ribosomal subunit interface upon signal sequence recognition.
The eukaryotic signal recognition particle (SRP) is essential for cotranslational targeting of proteins to the endoplasmic reticulum (ER). The SRP Alu domain is specifically required for delaying nascent chain elongation upon signal sequence recognition by SRP and was therefore proposed to interact directly with ribosomes. Using protein cross-linking, we provide experimental evidence that the Alu binding protein SRP14 is in close physical proximity of several ribosomal proteins in functional complexes. Cross-linking occurs even in the absence of a signal sequence in the nascent chain demonstrating that SRP can bind to all translating ribosomes and that close contacts between the Alu domain and the ribosome are independent of elongation arrest activity. Without a signal sequence, SRP14 cross-links predominantly to a protein of the large subunit. Upon signal sequence recognition, certain cross-linked products become detectable or more abundant revealing a change in the Alu domain-ribosome interface. At this stage, the Alu domain of SRP is located at the ribosomal subunit interface since SRP14 can be cross-linked to proteins from the large and small ribosomal subunits. Hence, these studies reveal differential modes of SRP-ribosome interactions mediated by the Alu domain.
Terzi
Lionel
L
Université de Genève, Département de Biologie Cellulaire, CH-1211 Genèva 4, Switzerland.
Pool
Martin R
MR
Dobberstein
Bernhard
B
Strub
Katharina
K
D020087:Alu Elements; D001665:Binding Sites; D017124:Conserved Sequence; D003432:Cross-Linking Reagents / Q000737:chemistry; D004268:DNA-Binding Proteins / Q000737:chemistry*; D008956:Models, Chemical; D008958:Models, Molecular; D021382:Protein Sorting Signals; D017434:Protein Structure, Tertiary; D021122:Protein Subunits / Q000737:chemistry*; D012261:Ribonucleoproteins / Q000737:chemistry*; D012269:Ribosomal Proteins / Q000737:chemistry*; D012270:Ribosomes / Q000737:chemistry; D018271:Signal Recognition Particle / Q000737:chemistry*
D003432:Cross-Linking Reagents; D004268:DNA-Binding Proteins; D021382:Protein Sorting Signals; D021122:Protein Subunits; D012261:Ribonucleoproteins; D012269:Ribosomal Proteins; D018271:Signal Recognition Particle