The Saccharomyces cerevisiae TIF6 gene encoding translation initiation factor 6 is required for 60S ribosomal subunit biogenesis

U. Basu, K. Si, J. R. Warner, U. Maitra

Research output: Contribution to journalArticle

117 Citations (Scopus)

Abstract

Eukaryotic translation initiation factor 6 (eIF6), a monomeric protein of about 26 kDa, can bind to the 60S ribosomal subunit and prevent its association with the 40S ribosomal subunit. In Saccharomyces cerevisiae, eIF6 is encoded by a single-copy essential gene. To understand the function of eIF6 in yeast cells, we constructed a conditional mutant haploid yeast strain in which a functional but a rapidly degradable form of eIF6 fusion protein was synthesized from a repressible GAL10 promoter. Depletion of eIF6 from yeast cells resulted in a selective reduction in the level of 60S ribosomal subunits, causing a stoichiometric imbalance in 60S-to-40S subunit ratio and inhibition of the rate of in vivo protein synthesis. Further analysis indicated that eIF6 is not required for the stability of 60S ribosomal subunits. Rather, eIF6-depleted cells showed defective pre-rRNA processing, resulting in accumulation of 35S pre-rRNA precursor, formation of a 23S aberrant pre-rRNA, decreased 20S pre-rRNA levels, and accumulation of 27SB pre-rRNA. The defect in the processing of 27S prerRNA resulted in the reduced formation of mature 25S and 5.8S rRNAs relative to 18S rRNA, which may account for the selective deficit of 60S ribosomal subunits in these cells. Cell fractionation as well as indirect immunofluorescence studies showed that c-Myc or hemagglutinin epitope-tagged eIF6 was distributed throughout the cytoplasm and the nuclei of yeast cells.

Original languageEnglish (US)
Pages (from-to)1453-1462
Number of pages10
JournalMolecular and Cellular Biology
Volume21
Issue number5
DOIs
StatePublished - 2001

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Eukaryotic Large Ribosome Subunits
Eukaryotic Initiation Factors
Peptide Initiation Factors
Saccharomyces cerevisiae
RNA Precursors
Genes
Yeasts
Eukaryotic Small Ribosome Subunits
Cell Fractionation
Proteins
eIF-6
Haploidy
Essential Genes
Hemagglutinins
Indirect Fluorescent Antibody Technique
Cell Nucleus
Epitopes
Cytoplasm

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics
  • Cell Biology

Cite this

The Saccharomyces cerevisiae TIF6 gene encoding translation initiation factor 6 is required for 60S ribosomal subunit biogenesis. / Basu, U.; Si, K.; Warner, J. R.; Maitra, U.

In: Molecular and Cellular Biology, Vol. 21, No. 5, 2001, p. 1453-1462.

Research output: Contribution to journalArticle

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