title: Interaction of the release factors with the Escherichia coli ribosome: structurally and functionally-important domains.
abstract: There are two major domains of interaction between the Escherichia coli release factors (RF-1 and RF-2) and each subunit of the ribosome. RF-2 has a binding domain on the shoulder and lower head region of the small subunit at the small lobe distant from the decoding site. This is in close proximity to one of the domains on the large subunit which includes the body dimer of L7/L12 and L11. The other domains of interaction, at the decoding site on the small subunit, and at the peptidyltransferase centre of the large subunit of the ribosome, are some distance from the first two, although the evidence for direct contact with the ribosome is less comprehensive. The release factors may therefore have two distinct structural domains, and in support of this concept RF-1 and RF-2 can both be cleaved into two fragments by papain. Region-specific antibodies, and antibodies against defined peptide within the RF sequences have given an indication that a significant part of an interacting RF molecule is in close proximity to the ribosome surface, confirming an observation by immunoelectron microscopy which suggested that the RF penetrates deeply into the cleft between the two subunits. A region of highly conserved primary sequence between the two release factors from E coli is also conserved in those from B subtilis suggesting it forms an important structural or functional domain. Antibodies against peptides from the N-terminal end of this region strongly inhibit binding of the RF to the ribosome.
authors.affiliation: Department of Biochemistry, University of Otago, Dunedin, New Zealand.
authors.forename: J G
authors.initials: JG
authors.lastname: Moffat
authors.forename: K M
authors.initials: KM
authors.lastname: Timms
authors.forename: C N
authors.initials: CN
authors.lastname: Trotman
authors.forename: W P
authors.initials: WP
authors.lastname: Tate
mesh_terms: D001665:Binding Sites; D004926:Escherichia coli / Q000378:metabolism; D007120:Immunochemistry; D015394:Molecular Structure; D010454:Peptide Termination Factors / Q000737:chemistry / Q000276:immunology / Q000378:metabolism*; D012270:Ribosomes / Q000378:metabolism* / Q000648:ultrastructure
chemical_list: D010454:Peptide Termination Factors; C041460:peptide chain termination release factor 2