[The use of cross-linking platinum reagents in the study of tRNA and mRNA interaction with ribosomes].
The use of some bifunctional Pt(II)-containing cross-linking reagents for investigation of structural organization of ribosomal tRNA- and mRNA-binding centres is demonstrated for various types of [70S ribosome.mRNA-tRNA] complexes. It is shown that treatment of the complexes [70S ribosome.Ac[14C]Phe-tRNA(Phe).poly(U)], [70S ribosome.3'-32pCp-tRNA(Phe).poly(U)] and [70S ribosome.f[35S]Met-tRNA(fMet).AUGU6] with Pt(II)-derivatives results in covalent attachment of tRNA to ribosome. AcPhe-tRNA(Phe) and 3'-pCp-tRNA(Phe) bound at the P site were found to be cross-linked preferentially to 30S subunit. fMet-tRNA(fMet) within the 70S initiation complex is cross-linked to both ribosome subunits approximately in the same extent, which exceeds two-fold the level of the tRNA(Phe) cross-linking. All used tRNA species were cross-linked in the comparable degree both to rRNA and proteins of both subunits in all types of the complexes studied. 32pAUGU6 cross-links exclusively to 30S subunit (to 16S RNA only) within [70S ribosome.32pAUGU6.fMet-tRNA(fMet)] complex. In the absence of fMet-tRNAfMet the level of the cross-linking is 4-fold lower.
Vladimirov
S N
SN
Graĭfer
D M
DM
Zenkova
M A
MA
Kazakov
S A
SA
Karpova
G G
GG
D055598:Chemical Phenomena; D002621:Chemistry; D003432:Cross-Linking Reagents; D009944:Organoplatinum Compounds; D010984:Platinum; D012333:RNA, Messenger / Q000378:metabolism*; D012343:RNA, Transfer / Q000378:metabolism*; D012270:Ribosomes / Q000378:metabolism*
D003432:Cross-Linking Reagents; D009944:Organoplatinum Compounds; D012333:RNA, Messenger; D010984:Platinum; D012343:RNA, Transfer
Primenenie sshivaiushchikh platinovykh reagentov dlia izucheniia vzaimodeĭstviia tRNK i mRNK s ribosomami.