Structure of the mammalian ribosome-channel complex at 17A resolution.
The co-translational translocation of proteins into the endoplasmic reticulum (ER) lumen and the biogenesis of membrane proteins require ribosome binding to a membrane channel formed by the Sec61p complex. We now report the 17A structure of a mammalian ribosome-channel complex derived from ER membranes. Atomic models of the ribosomal subunits were aligned to the programmed ribosome from Thermus thermophilus, to provide a common reference frame. The T.thermophilus ribosome, and by extension all known high resolution subunit models, were then docked within our map of the ribosome-channel complex. The structure shows that the ribosome contains a putative tRNA in the exit site, and a comparison with a non-programmed, yeast ribosome suggests that the L1 stalk may function as a gate in the tRNA exit path. We have localized six major expansion segments in the large subunit of the vertebrate ribosome including ES27, and suggest a function for ES30. The large ribosomal subunit is linked to the channel by four connections. We identified regions in the large subunit rRNA and four proteins that may help form the connections. These regions of the ribosome probably serve as a template to guide the assembly of the asymmetric translocation channel. Three of the connections form a picket fence that separates the putative translocation pore from the attachment site of an additional membrane component. The ribosome-channel connections also create an open junction that would allow egress of a nascent chain into the cytosol. At a threshold that is appropriate for the entire complex, the channel is rather solid and the lumenal half of the putative translocation pore is closed. These data suggest that the flow of small molecules across the membrane may be impeded by the channel itself, rather than the ribosome-channel junction.
Morgan
David Gene
DG
Department of Physiology and Biophysics, Boston University School of Medicine, 700 Albany St., Boston, MA 02118-2526, USA.
Ménétret
Jean-François
JF
Neuhof
Andrea
A
Rapoport
Tom A
TA
Akey
Christopher W
CW
D000818:Animals; D020285:Cryoelectron Microscopy; D004285:Dogs; D018870:Endoplasmic Reticulum, Rough / Q000737:chemistry* / Q000378:metabolism / Q000648:ultrastructure; D007425:Intracellular Membranes / Q000737:chemistry / Q000378:metabolism / Q000648:ultrastructure; D008565:Membrane Proteins / Q000737:chemistry / Q000378:metabolism; D008958:Models, Molecular; D014176:Protein Biosynthesis; D011487:Protein Conformation; D011500:Protein Synthesis Inhibitors / Q000494:pharmacology; D021381:Protein Transport; D011691:Puromycin / Q000494:pharmacology; D012343:RNA, Transfer / Q000378:metabolism; D012270:Ribosomes / Q000737:chemistry* / Q000378:metabolism / Q000648:ultrastructure*; D012492:Salts; D016415:Sequence Alignment; D016963:Thermus thermophilus / Q000737:chemistry
D008565:Membrane Proteins; D011500:Protein Synthesis Inhibitors; D012492:Salts; D011691:Puromycin; D012343:RNA, Transfer