Protein synthesis in rabbit reticulocytes: Requirements for Met-tRNAf·40S preinitiation complex formation with AUG-codon and physiological mRNAs

Reena Roy, Nargis Nasrin, Mir Firoz Ahmad, Naba K. Gupta

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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.

Original languageEnglish (US)
Pages (from-to)1418-1425
Number of pages8
JournalBiochemical and Biophysical Research Communications
Volume122
Issue number3
DOIs
Publication statusPublished - Aug 16 1984

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All Science Journal Classification (ASJC) codes

  • Biophysics
  • Biochemistry
  • Molecular Biology
  • Cell Biology

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