[Interaction of deacylated phenylalanyl tRNA from yeasts with Escherichia coli ribosomes. The role of the modified nucleotide in codon-anticodon interaction].
The method of anticodon loop replacement has been used to make derivatives of yeast tRNA(Phe)GmAAY with the substitution at the 37 position (tRNA(Phe)GAAA), and at both the anticodon (tRNA(Phe)GCAG) and the 37 position. A quantitative study of the interaction of various types of yeast deacylated tRNA: tRNA(Phe)GmAAY, tRNA(Phe)GAAA, tRNA(Phe)GCAG, and tRNA(Phe)-Y with the P site of the 70S ribosome.poly(U) complex was carried out at different Mg2+ concentrations and temperatures. The replacement of the Y base on the nonmodified adenosine decreases the interaction enthalpy from 39 to 24 kcal/mole, whereas the complete removal of the Y base reduces the interaction enthalpy to 16 kcal/mole. The replacement of the second letter of the anticodon (A) with cytosine leads to a drop in the enthalpy to 6 kcal/mole, which is typical of tRNA interaction with the P site in the absence of poly(U). In the absence of poly(U) the affinity of tRNA(Phe)-Y for the P site of the 70S ribosome is 5 times lower than the affinity of tRNA(Phe)GmAAY and tRNA(Phe)GCAG. Thus, in the ribosome the modified nucleotide not only stabilizes the codon-anticodon interaction owing to the stacking interaction with the stack of codon-anticodon bases, but also lowers the free energy of binding as a result of the interaction of the modified nucleotide itself with the hydrophobic center of the P site on the ribosome.
Katunin
V I
VI
Soboleva
N G
NG
Makhno
V I
VI
Sedel'nikova
E A
EA
Zhenodarova
S M
SM
Kirillov
S V
SV
D000215:Acylation; D000926:Anticodon; D003062:Codon; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D009711:Nucleotides / Q000737:chemistry / Q000378:metabolism*; D012360:RNA, Transfer, Phe / Q000378:metabolism*; D012270:Ribosomes / Q000378:metabolism*; D012441:Saccharomyces cerevisiae / Q000235:genetics*; D013816:Thermodynamics
D000926:Anticodon; D003062:Codon; D009711:Nucleotides; D012360:RNA, Transfer, Phe
Vzaimodeĭstvie deatsilirovannoĭ fenilalaninovoĭ tRNK iz drozhzheĭ s ribosomami Escherichia coli. Rol' modifitsirovannogo nukleotida v kodon-antikodonovom vazimodeĭstvii.