Related eIF3 subunits TIF32 and HCR1 interact with an RNA recognition motif in PRT1 required for eIF3 integrity and ribosome binding.
eIF3 binds to 40S ribosomal subunits and stimulates recruitment of Met-tRNAiMet and mRNA to the pre-initiation complex. Saccharomyces cerevisiae contains an ortholog of human eIF3 subunit p35, HCR1, whose interactions with yeast eIF3 are not well defined. We found that HCR1 has a dual function in translation initiation: it binds to, and stabilizes, the eIF3-eIF5- eIF1-eIF2 multifactor complex and is required for the normal level of 40S ribosomes. The RNA recognition motif (RRM) of eIF3 subunit PRT1 interacted simultaneously with HCR1 and with an internal domain of eIF3 subunit TIF32 that has sequence and functional similarity to HCR1. PRT1, HCR1 and TIF32 were also functionally linked by genetic suppressor analysis. We propose that HCR1 stabilizes or modulates interaction between TIF32 and the PRT1 RRM. Removal of the PRT1 RRM resulted in dissociation of TIF32, NIP1, HCR1 and eIF5 from eIF3 in vivo, and destroyed 40S ribosome binding by the residual PRT1-TIF34-TIF35 subcomplex. Hence, the PRT1 RRM is crucial for the integrity and ribosome-binding activity of eIF3.
Valásek
L
L
Laboratory of Eukaryotic Gene Regulation, National Institute of Child Health and Human Development, Bethesda, MD 20892, USA.
Phan
L
L
Schoenfeld
L W
LW
Valásková
V
V
Hinnebusch
A G
AG
D000483:Alleles; D000595:Amino Acid Sequence; D018797:Cell Cycle Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D039621:Eukaryotic Initiation Factor-3; D005656:Fungal Proteins / Q000737:chemistry / Q000378:metabolism*; D016153:Genes, Suppressor; D008969:Molecular Sequence Data; D009154:Mutation; D010448:Peptide Initiation Factors / Q000235:genetics / Q000378:metabolism*; D039665:Prokaryotic Initiation Factor-3; D011485:Protein Binding; D014176:Protein Biosynthesis; D012331:RNA, Fungal / Q000378:metabolism*; D012270:Ribosomes / Q000378:metabolism*; D012441:Saccharomyces cerevisiae / Q000235:genetics / Q000378:metabolism; D029701:Saccharomyces cerevisiae Proteins; D017386:Sequence Homology, Amino Acid
D018797:Cell Cycle Proteins; D039621:Eukaryotic Initiation Factor-3; D005656:Fungal Proteins; C121939:HCR1 protein, S cerevisiae; D010448:Peptide Initiation Factors; D039665:Prokaryotic Initiation Factor-3; D012331:RNA, Fungal; C111889:RPG1 protein, S cerevisiae; D029701:Saccharomyces cerevisiae Proteins