title: The phenotype of many independently isolated +1 frameshift suppressor mutants supports a pivotal role of the P-site in reading frame maintenance.
abstract: The main features of translation are similar in all organisms on this planet and one important feature of it is the way the ribosome maintain the reading frame. We have earlier characterized several bacterial mutants defective in tRNA maturation and found that some of them correct a +1 frameshift mutation; i.e. such mutants possess an error in reading frame maintenance. Based on the analysis of the frameshifting phenotype of such mutants we proposed a pivotal role of the ribosomal grip of the peptidyl-tRNA to maintain the correct reading frame. To test the model in an unbiased way we first isolated many (467) independent mutants able to correct a +1 frameshift mutation and thereafter tested whether or not their frameshifting phenotypes were consistent with the model. These 467+1 frameshift suppressor mutants had alterations in 16 different loci of which 15 induced a defective tRNA by hypo- or hypermodifications or altering its primary sequence. All these alterations of tRNAs induce a frameshift error in the P-site to correct a +1 frameshift mutation consistent with the proposed model. Modifications next to and 3' of the anticodon (position 37), like 1-methylguanosine, are important for proper reading frame maintenance due to their interactions with components of the ribosomal P-site. Interestingly, two mutants had a defect in a locus (rpsI), which encodes ribosomal protein S9. The C-terminal of this protein contacts position 32-34 of the peptidyl-tRNA and is thus part of the P-site environment. The two rpsI mutants had a C-terminal truncated ribosomal protein S9 that destroys its interaction with the peptidyl-tRNA resulting in +1 shift in the reading frame. The isolation and characterization of the S9 mutants gave strong support of our model that the ribosomal grip of the peptidyl-tRNA is pivotal for the reading frame maintenance.
authors.affiliation: Department of Molecular Biology, Umeå University, Umeå, Sweden.
authors.forename: Gunilla
authors.initials: G
authors.lastname: Jäger
authors.forename: Kristina
authors.initials: K
authors.lastname: Nilsson
authors.forename: Glenn R
authors.initials: GR
authors.lastname: Björk
mesh_terms: D000595:Amino Acid Sequence; D000926:Anticodon; D001426:Bacterial Proteins / Q000737:chemistry / Q000235:genetics; D001483:Base Sequence; D003062:Codon; D016368:Frameshift Mutation; D008969:Molecular Sequence Data; D009690:Nucleic Acid Conformation; D009876:Operon / Q000502:physiology; D010641:Phenotype; D014176:Protein Biosynthesis / Q000502:physiology; D012346:RNA, Transfer, Amino Acyl / Q000737:chemistry / Q000502:physiology*; D016364:Reading Frames; D000091243:Ribosomal Protein S9; D012269:Ribosomal Proteins / Q000737:chemistry; D012270:Ribosomes / Q000502:physiology*
chemical_list: D000926:Anticodon; D001426:Bacterial Proteins; D003062:Codon; D012346:RNA, Transfer, Amino Acyl; D000091243:Ribosomal Protein S9; D012269:Ribosomal Proteins; C019037:tRNA, peptidyl-