title: Understanding ribosome assembly: the structure of in vivo assembled immature 30S subunits revealed by cryo-electron microscopy.
abstract: Four decades after early in vitro assembly studies demonstrated that ribosome assembly is a controlled process, our understanding of ribosome assembly is still incomplete. Just as structure determination has been so important to understanding ribosome function, so too will it be critical to sorting out the assembly process. Here, we used a viable deletion in the yjeQ gene, a recognized ribosome assembly factor, to isolate and structurally characterize immature 30S subunits assembled in vivo. These small ribosome subunits contained unprocessed 17S rRNA and lacked some late ribosomal proteins. Cryo-electron microscopy reconstructions revealed that the presence of precursor sequences in the rRNA induces a severe distortion in the 3' minor domain of the subunit involved in the decoding of mRNA and interaction with the large ribosome subunit. These findings suggest that rRNA processing events induce key local conformational changes directing the structure toward the mature assembly. We concluded that rRNA processing, folding, and the entry of tertiary r-proteins are interdependent events in the late stages of 30S subunit assembly. In addition, we demonstrate how studies of emerging assembly factors in ribosome biogenesis can help to elucidate the path of subunit assembly in vivo.
authors.affiliation: Department of Biochemistry and Biomedical Sciences, McMaster University, Hamilton, Ontario L8N3Z5, Canada.
authors.forename: Ahmad
authors.initials: A
authors.lastname: Jomaa
authors.forename: Geordie
authors.initials: G
authors.lastname: Stewart
authors.forename: Jaime
authors.initials: J
authors.lastname: Martín-Benito
authors.forename: Ryszard
authors.initials: R
authors.lastname: Zielke
authors.forename: Tracey L
authors.initials: TL
authors.lastname: Campbell
authors.forename: Janine R
authors.initials: JR
authors.lastname: Maddock
authors.forename: Eric D
authors.initials: ED
authors.lastname: Brown
authors.forename: Joaquin
authors.initials: J
authors.lastname: Ortega
mesh_terms: D020285:Cryoelectron Microscopy; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism*; D029968:Escherichia coli Proteins / Q000235:genetics; D020558:GTP Phosphohydrolases / Q000235:genetics; D017353:Gene Deletion; D017433:Protein Structure, Secondary; D012335:RNA, Ribosomal / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D054680:Ribosome Subunits, Small, Bacterial / Q000737:chemistry / Q000378:metabolism* / Q000648:ultrastructure
chemical_list: D029968:Escherichia coli Proteins; D012335:RNA, Ribosomal; D020558:GTP Phosphohydrolases; C467318:RsgA protein, E coli