Reconstitution of mammalian 48S ribosomal translation initiation complex Journal Article
| Authors: | Majumdar, R.; Chaudhuri, J.; Maitra, U. |
| Article Title: | Reconstitution of mammalian 48S ribosomal translation initiation complex |
| 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. © 2007 Elsevier Inc. All rights reserved. |
| Keywords: | genetics; review; translation initiation; animal; metabolism; mammalia; animals; chemistry; messenger rna; protein synthesis; rna caps; eukaryota; mammal; recombinant proteins; recombinant protein; protein biosynthesis; adenosine triphosphate; cell free system; guanosine triphosphate; eukaryotic initiation factors; ribosomes; start codon; codon, initiator; protein rna binding; capped rna; rabbits; ribosome; initiation factor; rabbit; cell-free system; ribosome subunit; methionine transfer rna; 40s ribosomal subunit; 48s ribosomal subunit; 60s ribosomal subunit; 80s ribosomal subunit; rna, transfer, met |
| Journal Title: | Methods in Enzymology |
| Volume: | 430 |
| ISSN: | 0076-6879 |
| Publisher: | Academic Press |
| Date Published: | 2007-01-01 |
| Start Page: | 179 |
| End Page: | 208 |
| Language: | English |
| DOI: | 10.1016/s0076-6879(07)30008-6 |
| PUBMED: | 17913639 |
| PROVIDER: | scopus |
| DOI/URL: | |
| Notes: | Chapter 8 in "Translation Initiation: Reconstituted Systems and Biophysical Methods" (ISBN: 978-0-12-373969-8) - Export Date: 17 November 2011 - CODEN: MENZA - Source: Scopus |
