Reconstitution of Mammalian 48S Ribosomal Translation Initiation Complex

Romit Majumdar, Jayanta Chaudhuri, Umadas Maitra

Research output: Contribution to journalArticle

1 Citation (Scopus)

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)
Pages (from-to)179-208
Number of pages30
JournalMethods in Enzymology
Volume430
DOIs
StatePublished - 2007

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Eukaryotic Small Ribosome Subunits
RNA, Transfer, Met
Messenger RNA
Initiator Codon
Eukaryotic Large Ribosome Subunits
Eukaryotic Initiation Factors
Guanosine Triphosphate
Ribosomes
Elongation
Adenosine Triphosphate
Scanning
Molecules

ASJC Scopus subject areas

  • Biochemistry

Cite this

Reconstitution of Mammalian 48S Ribosomal Translation Initiation Complex. / Majumdar, Romit; Chaudhuri, Jayanta; Maitra, Umadas.

In: Methods in Enzymology, Vol. 430, 2007, p. 179-208.

Research output: Contribution to journalArticle

Majumdar, Romit ; Chaudhuri, Jayanta ; Maitra, Umadas. / Reconstitution of Mammalian 48S Ribosomal Translation Initiation Complex. In: Methods in Enzymology. 2007 ; Vol. 430. pp. 179-208.
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