The ribosomal grip of the peptidyl-tRNA is critical for reading frame maintenance.
If a ribosome shifts to an alternative reading frame during translation, the information in the message is usually lost. We have selected mutants of Salmonella typhimurium with alterations in tRNA(cmo5UGG)(Pro) that cause increased frameshifting when present in the ribosomal P-site. In 108 such mutants, two parts of the tRNA molecule are altered: the anticodon stem and the D-arm, including its tertiary interactions with the variable arm. Some of these alterations in tRNA(cmo5UGG)(Pro) are in close proximity to ribosomal components in the P-site. The crystal structure of the 30S subunit suggests that the C-terminal end of ribosomal protein S9 contacts nucleotides 32-34 of peptidyl-tRNA. We have isolated mutants with defects in the C-terminus of S9 that induce +1 frameshifting. Combinations of changes in tRNA(cmo5UGG)(Pro) and S9 suggest that an interaction occurs between position 32 of the peptidyl-tRNA and the C-terminal end of S9. Together, our results suggest that the cause of frameshifting is an aberrant interaction between the peptidyl-tRNA and the P-site environment. We suggest that the "ribosomal grip" of the peptidyl-tRNA is pivotal for maintaining the reading frame.
Näsvall
S Joakim
SJ
Department of Molecular Biology, Umeå University, S-901 87 Umeå, Sweden.
Nilsson
Kristina
K
Björk
Glenn R
GR
D018965:Frameshifting, Ribosomal; D008958:Models, Molecular; D020125:Mutation, Missense; D009690:Nucleic Acid Conformation; D017354:Point Mutation; D014176:Protein Biosynthesis; D012333:RNA, Messenger / Q000378:metabolism*; D012346:RNA, Transfer, Amino Acyl / Q000235:genetics / Q000378:metabolism*; D012361:RNA, Transfer, Pro / Q000235:genetics / Q000378:metabolism; D016364:Reading Frames; D000091243:Ribosomal Protein S9; D012269:Ribosomal Proteins / Q000235:genetics; D012270:Ribosomes / Q000378:metabolism*; D012486:Salmonella typhimurium / Q000502:physiology*
D012333:RNA, Messenger; D012346:RNA, Transfer, Amino Acyl; D012361:RNA, Transfer, Pro; D000091243:Ribosomal Protein S9; D012269:Ribosomal Proteins; C019037:tRNA, peptidyl-