Reconstitution of Mammalian 48S Ribosomal Translation Initiation Complex

Romit Majumdar, Jayanta Chaudhuri, Umadas Maitra

Research output: Chapter in Book/Report/Conference proceedingChapter

2 Scopus citations

Abstract

Initiation of translation is defined as the process by which a 40S ribosomal subunit, containing bound initiator methionyl-tRNA (Met-tRNAi), is positioned at the initiation AUG codon of an mRNA to form the 48S initiation complex. Subsequently, a 60S ribosomal subunit joins the 48S initiation complex to form an elongation-competent 80S initiation complex. By use of highly purified eukaryotic translation initiation factors (eIFs), ribosomes, Met-tRNAi, mRNA, GTP as an effector molecule, and ATP as a source of energy, the initiation step of translation can be efficiently reconstituted. In this chapter, we describe the detailed procedure for efficient binding of Met-tRNAi to the 40S ribosomal subunit, the subsequent binding of the resulting 43S preinitiation complex to an mRNA, and scanning and positioning of the 43S complex at the AUG start codon of the mRNA to form the 48S initiation complex.

Original languageEnglish (US)
Title of host publicationTranslation Initiation
Subtitle of host publicationReconstituted Systems and Biophysical Methods
PublisherAcademic Press Inc.
Pages179-208
Number of pages30
ISBN (Print)9780123739698
DOIs
StatePublished - Jan 1 2007

Publication series

NameMethods in Enzymology
Volume430
ISSN (Print)0076-6879

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology

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    Majumdar, R., Chaudhuri, J., & Maitra, U. (2007). Reconstitution of Mammalian 48S Ribosomal Translation Initiation Complex. In Translation Initiation: Reconstituted Systems and Biophysical Methods (pp. 179-208). (Methods in Enzymology; Vol. 430). Academic Press Inc.. https://doi.org/10.1016/S0076-6879(07)30008-6