[Translocation in ribosomes induced by elongation factor G without cleavage of GTP. Study using a solid phase translation system in columns].
Columns containing ribosomes translating poly(U) covalently bound with cellulose (solid-phase translating system) were used to study translocation in ribosomes. It is shown that the passing of elongation factor G (EF-G) with the non-cleavable analog of GTP (GMP-PCP) through a column containing pre-translocated ribosomes results in the increase of competence for puromycin (i. e. to the transition of pre-translocated peptidyl-tRNA into the post-translocated state) just as in the case of the passing of EF-G with GTP. On the other hand, it is shown that the passing of EF-G with GMP-PCP through a column with pre-translocated ribosomes makes them capable of binding the next aminoacyl-tRNA (i. e. leads to the vacation of the ribosomal A-site). Thus, by means of the two independent tests it is shown that EF-G-promoted translocation in the ribosome can proceed without GTP hydrolysis. On the basis of the data obtained, a controlled step-wise elongation of polypeptide with the participation of EF-G without GTP cleavage has been carried out in the solid-phase column system of translation.
D006160:Guanosine Triphosphate / Q000378:metabolism*; D010441:Peptide Chain Elongation, Translational; D010445:Peptide Elongation Factors / Q000494:pharmacology*; D011072:Poly U / Q000378:metabolism; D014176:Protein Biosynthesis; D011691:Puromycin / Q000494:pharmacology; D012346:RNA, Transfer, Amino Acyl / Q000378:metabolism; D012270:Ribosomes / Q000378:metabolism*
D010445:Peptide Elongation Factors; D012346:RNA, Transfer, Amino Acyl; D011072:Poly U; D011691:Puromycin; D006160:Guanosine Triphosphate
Translokatsiia v ribosomakh, indutsiruemaia faktorom élongatsii G bez raspada GTP. Izuchenie pri pomoshchi tverdofaznoĭ sistemy transliatsii v kolonkakh.