Topography of the Escherichia coli initiation factor 2/fMet-tRNA(f)(Met) complex as studied by cross-linking.
trans-Diamminedichloroplatinum(II) was used to induce reversible cross-links between Escherichia coli initiation factor 2 (IF-2) and fMet-tRNA(f)(Met). Two distinct cross-links between IF-2 and the initiator tRNA were produced. Analysis of the cross-linking regions on both RNA and protein moieties reveals that the T arm of the tRNA is in the proximity of a region of the C-terminal domain of IF-2 (residues Asn611-Arg645). This cross-link is well-correlated with the fact that the C-domain of IF-2 contains the fMet-tRNA binding site and that the cross-linked RNA fragment precisely maps in a region which is protected by IF-2 from chemical modification and enzymatic digestion. Rather unexpectedly, a second cross-link was characterized which involves the anticodon arm of fMet-tRNA(f)(Met) and the N-terminal part of IF-2 (residues Trp215-Arg237).
Yusupova
G
G
Institut de Biologie Moléculaire et Cellulaire, Strasbourg, France.
Reinbolt
J
J
Wakao
H
H
Laalami
S
S
Grunberg-Manago
M
M
Romby
P
P
Ehresmann
B
B
Ehresmann
C
C
D000595:Amino Acid Sequence; D001483:Base Sequence; D002945:Cisplatin / Q000494:pharmacology*; D003432:Cross-Linking Reagents; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli / Q000187:drug effects / Q000378:metabolism*; D015852:Eukaryotic Initiation Factor-2 / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism*; D007700:Kinetics; D008969:Molecular Sequence Data; D009690:Nucleic Acid Conformation; D010446:Peptide Fragments / Q000737:chemistry / Q000302:isolation & purification; D011487:Protein Conformation; D012358:RNA, Transfer, Met / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism*; D013379:Substrate Specificity
D003432:Cross-Linking Reagents; D015852:Eukaryotic Initiation Factor-2; D010446:Peptide Fragments; D012358:RNA, Transfer, Met; C089238:fMet-tRNA(fMet); C034697:transplatin; D002945:Cisplatin