GTP binding and hydrolysis by the signal recognition particle during initiation of protein translocation.
The signal recognition particle (SRP) consists of one RNA and six protein subunits. The N-terminal domain of the 54K subunit contains a putative GTP-binding site, whereas the C-terminal domain binds signal sequences and SRP RNA. Binding of SRP to the signal sequence as it emerges from the ribosome creates a cytosolic targeting complex containing the nascent polypeptide chain, the translating ribosome, and SRP. This complex is directed to the endoplasmic reticulum membrane as a result of its interaction with the SRP receptor, a membrane protein composed of two subunits, SR alpha and SR beta, each of which also contains a GTP-binding domain. In the presence of GTP, SRP receptor binding to SRP causes the latter to dissociate from both the signal sequence and the ribosome. GTP is then hydrolysed so that SRP can be released from the SRP receptor and returned to the cytosol. Here we show that the 54K subunit (M(r) 54,000) of SRP (SRP54) is a GTP-binding protein stabilized in a nucleotide-free state by signal sequences, and that the SRP receptor both increases the affinity of SRP54 for GTP and activates its GTPase. We propose that nucleotide-mediated conformational changes in SRP54 regulate the release of signal sequences and the docking of ribosomes at the endoplasmic reticulum.
Miller
J D
JD
Department of Biochemistry and Biophysics, University of California Medical School, San Francisco 94143-0448.
Wilhelm
H
H
Gierasch
L
L
Gilmore
R
R
Walter
P
P
D000595:Amino Acid Sequence; D001692:Biological Transport / Q000502:physiology; D004721:Endoplasmic Reticulum / Q000378:metabolism; D004926:Escherichia coli / Q000378:metabolism; D019204:GTP-Binding Proteins / Q000378:metabolism*; D006160:Guanosine Triphosphate / Q000378:metabolism*; D006868:Hydrolysis; D008969:Molecular Sequence Data; D010446:Peptide Fragments / Q000378:metabolism; D011499:Protein Processing, Post-Translational / Q000502:physiology*; D018160:Receptors, Cytoplasmic and Nuclear / Q000378:metabolism; D018000:Receptors, Peptide / Q000378:metabolism; D018271:Signal Recognition Particle / Q000378:metabolism*
D010446:Peptide Fragments; D018160:Receptors, Cytoplasmic and Nuclear; D018000:Receptors, Peptide; D018271:Signal Recognition Particle; C036318:signal peptide receptor; D006160:Guanosine Triphosphate; D019204:GTP-Binding Proteins