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Protein synthesis in rabbit reticulocytes: requirements for Met-tRNAf . 40S preinitiation complex formation with AUG-codon and physiological mRNAs |
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| Authors: | R Roy N Nasrin M F Ahmad N K Gupta |
| Institution: | Department of Chemistry, University of Nebraska Lincoln, Nebraska 68588-0304 USA |
| Abstract: | Under standard conditions, in the presence of GTP, highly purified eIF-2 and Co-eIF-2 factor preparations efficiently stimulated AUG-codon dependent but not physiological mRNA-dependent Met-tRNAf binding to 40S ribosomes. Replacement of GTP by a nonhydrolyzable GTP analog, GMP-PNP, in the above system, gave significant stimulation of Met-tRNAf binding to 40S ribosomes dependent on physiological mRNAs. Lower but significant stimulation of Met-tRNAf binding to 40S ribosomes was also observed when GTP was used in the presence of nucleoside 5'-diphosphate kinase (NDK) and ATP. ATP alone in the absence of NDK had no significant effect. This is the first report on the formation of a stable Met-tRNAf . 40S initiation complex dependent on physiological mRNAs and the factor requirements for such complex formation. |
| Keywords: | eIF-2 Co-eIF-2 (a high molecular weight protein complex) stimulates ternary complex formation by eIF-2 and also promotes GDP displacement from eIF-2·GDP during ternary complex formation NDK nucleoside 5′-diphosphate kinase GMP-PNP 5′-guanylyl-β γ-imidodiphosphate |
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