title: Adenine and guanine recognition of stop codon is mediated by different N domain conformations of translation termination factor eRF1.
abstract: Positioning of release factor eRF1 toward adenines and the ribose-phosphate backbone of the UAAA stop signal in the ribosomal decoding site was studied using messenger RNA (mRNA) analogs containing stop signal UAA/UAAA and a photoactivatable cross-linker at definite locations. The human eRF1 peptides cross-linked to these analogs were identified. Cross-linkers on the adenines at the 2nd, 3rd or 4th position modified eRF1 near the conserved YxCxxxF loop (positions 125-131 in the N domain), but cross-linker at the 4th position mainly modified the tripeptide 26-AAR-28. This tripeptide cross-linked also with derivatized 3'-phosphate of UAA, while the same cross-linker at the 3'-phosphate of UAAA modified both the 26-28 and 67-73 fragments. A comparison of the results with those obtained earlier with mRNA analogs bearing a similar cross-linker at the guanines indicates that positioning of eRF1 toward adenines and guanines of stop signals in the 80S termination complex is different. Molecular modeling of eRF1 in the 80S termination complex showed that eRF1 fragments neighboring guanines and adenines of stop signals are compatible with different N domain conformations of eRF1. These conformations vary by positioning of stop signal purines toward the universally conserved dipeptide 31-GT-32, which neighbors guanines but is oriented more distantly from adenines.
authors: Institute of Chemical Biology and Fundamental Medicine, Siberian Branch of the Russian Academy of Sciences, Novosibirsk, 630090, Russia.
authors: Konstantin N
authors: KN
authors: Bulygin
authors: Yulia S
authors: YS
authors: Khairulina
authors: Petr M
authors: PM
authors: Kolosov
authors: Aliya G
authors: AG
authors: Ven'yaminova
authors: Dmitri M
authors: DM
authors: Graifer
authors: Yuri N
authors: YN
authors: Vorobjev
authors: Ludmila Yu
authors: LY
authors: Frolova
authors: Galina G
authors: GG
authors: Karpova
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