Structural arrangement of tRNA binding sites on Escherichia coli ribosomes, as revealed from data on affinity labelling with photoactivatable tRNA derivatives.
A systematic study of protein environment of tRNA in ribosomes in model complexes representing different translation steps was carried out using the affinity labelling of the ribosomes with tRNA derivatives bearing aryl azide groups scattered statistically over tRNA guanine residues. Analysis of the proteins crosslinked to tRNA derivatives showed that the location of the derivatives in the aminoacyl (A) site led to the labelling of the proteins S5 and S7 in all complexes studied, whereas the labelling of the proteins S2, S8, S9, S11, S14, S16, S17, S18, S19, S21 as well as L9, L11, L14, L15, L21, L23, L24, L29 depended on the state of tRNA in A site. Similarly, the location of tRNA derivatives in the peptidyl (P) site resulted in the labelling of the proteins L27, S11, S13 and S19 in all states, whereas the labelling of the proteins S5, S7, S9, S12, S14, S20, S21 as well as L2, L13, L14, L17, L24, L27, L31, L32, L33 depended on the type of complex. The derivatives of tRNA(fMet) were found to crosslink to S1, S3, S5, S7, S9, S14 and L1, L2, L7/L12, L27. Based on the data obtained, a general principle of the dynamic functioning of ribosomes has been proposed: (i) the formation of each type of ribosomal complex is accompanied by changes in mutual arrangement of proteins - 'conformational adjustment' of the ribosome - and (ii) a ribosome can dynamically change its internal structure at each step of initiation and elongation; on the 70 S ribosome there are no rigidly fixed structures forming tRNA-binding sites (primarily A and P sites).
Graifer
D M
DM
Institute of Bioorganic Chemistry, Siberian Division of the Academy of Sciences of the U.S.S.R., Novosibirsk.
Babkina
G T
GT
Matasova
N B
NB
Vladimirov
S N
SN
Karpova
G G
GG
Vlassov
V V
VV
D000345:Affinity Labels; D001665:Binding Sites; D003432:Cross-Linking Reagents; D004139:Dinitrofluorobenzene / Q000031:analogs & derivatives; D004926:Escherichia coli / Q000235:genetics*; D009150:Mustard Compounds; D010777:Photochemistry; D011487:Protein Conformation; D012343:RNA, Transfer / Q000031:analogs & derivatives / Q000378:metabolism*; D012269:Ribosomal Proteins / Q000378:metabolism; D012270:Ribosomes / Q000378:metabolism*
D000345:Affinity Labels; D003432:Cross-Linking Reagents; D009150:Mustard Compounds; D012269:Ribosomal Proteins; C025881:4(N)-(2-chloroethyl-N-methylamino)benzylamine; C025880:2,4-dinitro-5-fluorophenylazide; D012343:RNA, Transfer; D004139:Dinitrofluorobenzene