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PMID: 9671501 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Identification of a translation initiation factor 3 (eIF3) core complex, conserved in yeast and mammals, that interacts with eIF5.

Molecular and cellular biology ·Vol. 18 ·No. 8 ·1998-08-00 ·Pages 4935-46

Phan L, Zhang X, Asano K, Anderson J, Vornlocher HP, Greenberg JR, Qin J, Hinnebusch AG

Abstract

Only five of the nine subunits of human eukaryotic translation initiation factor 3 (eIF3) have recognizable homologs encoded in the Saccharomyces cerevisiae genome, and only two of these (Prt1p and Tif34p) were identified previously as subunits of yeast eIF3. We purified a polyhistidine-tagged form of Prt1p (His-Prt1p) by Ni2+ affinity and gel filtration chromatography and obtained a complex of approximately 600 kDa composed of six polypeptides whose copurification was completely dependent on the polyhistidine tag on His-Prt1p. All five polypeptides associated with His-Prt1p were identified by mass spectrometry, and four were found to be the other putative homologs of human eIF3 subunits encoded in S. cerevisiae: YBR079c/Tif32p, Nip1p, Tif34p, and YDR429c/Tif35p. The fifth Prt1p-associated protein was eIF5, an initiation factor not previously known to interact with eIF3. The purified complex could rescue Met-tRNAiMet binding to 40S ribosomes in defective extracts from a prt1 mutant or extracts from which Nip1p had been depleted, indicating that it possesses a known biochemical activity of eIF3. These findings suggest that Tif32p, Nip1p, Prt1p, Tif34p, and Tif35p comprise an eIF3 core complex, conserved between yeast and mammals, that stably interacts with eIF5. Nip1p bound to eIF5 in yeast two-hybrid and in vitro protein binding assays. Interestingly, Sui1p also interacts with Nip1p, and both eIF5 and Sui1p have been implicated in accurate recognition of the AUG start codon. Thus, eIF5 and Sui1p may be recruited to the 40S ribosomes through physical interactions with the Nip1p subunit of eIF3.

MeSH Terms
Animals Epitopes Eukaryotic Initiation Factor-1 Eukaryotic Initiation Factor-3 Eukaryotic Initiation Factor-4G Eukaryotic Initiation Factor-5 Fungal Proteins/isolation & purification,metabolism Histidine Humans Mammals Mass Spectrometry Molecular Weight Nuclear Proteins/metabolism Peptide Initiation Factors/metabolism Peptides Precipitin Tests RNA, Transfer, Met/metabolism Ribosomes/metabolism Saccharomyces cerevisiae Saccharomyces cerevisiae Proteins Transcription Factors/metabolism
Chemicals
Epitopes Eukaryotic Initiation Factor-1 Eukaryotic Initiation Factor-3 Eukaryotic Initiation Factor-4G Eukaryotic Initiation Factor-5 Fungal Proteins Met-tRNA(i)(Met) NIP1 protein, S cerevisiae Nuclear Proteins Peptide Initiation Factors Peptides Prt1 protein, S cerevisiae RNA, Transfer, Met SUI1 protein, S cerevisiae Saccharomyces cerevisiae Proteins TIF34 protein, S cerevisiae Transcription Factors polyhistidine Histidine
Authors & Affiliations
8 authors, click to expand affiliations / ORCID
Phan L
Laboratory of Eukaryotic Gene Regulation, National Institute of Child Health and Human Development, Bethesda, Maryland 20892, USA.
Zhang X
Asano K
Anderson J
Vornlocher H P
Greenberg J R
Qin J
Hinnebusch A G
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Article Info
Journal
Molecular and cellular biology
Abbr.
Mol Cell Biol
ISSN
0270-7306
DOI
10.1128/MCB.18.8.4935
Published
1998-08-00
Pages
4935-46
Language
English
Region
United States
NLM ID
8109087
PMCID
PMC109077
Subset
IM
External Links
PubMed source DOI: 10.1128/MCB.18.8.4935 PMC Full Text View journal details
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