Saccharomyces cerevisiae protein Pci8p and human protein eIF3e/Int-6 interact with the eIF3 core complex by binding to cognate eIF3b subunits.
Mammalian, plant, and Schizosaccharomyces pombe eukaryotic initiation factor-3 (eIF3) contains a protein homologous to the product of int-6 (eIF3e), a frequent integration site of mouse mammary tumor viruses. By contrast, Saccharomyces cerevisiae does not encode a protein closely related to eIF3e/Int-6. Here, we characterize a novel S. cerevisiae protein (Pci8p, Yil071cp) that contains a PCI (proteasome-COP9 signalosome-eIF3) domain conserved in eIF3e/Int-6. We show that both Pci8p and human eIF3e/Int-6 expressed in budding yeast interact with the yeast eIF3 complex in vivo and in vitro by binding to a discrete segment of its eIF3b subunit Prt1p and that human eIF3e/Int-6 interacts with the human eIF3b segment homologous to the Pci8p-binding site of yeast Prt1p. These results refine our understanding of subunit interactions in the eIF3 complex and suggest structural similarity between human eIF3e/Int-6 and yeast Pci8p. However, deletion of PCI8 had no discernible effect on cell growth or translation initiation as judged by polysome analysis, suggesting that Pci8p is not required for the essential function of eIF3 in translation initiation. Motivated by the involvement of Int-6 in transcriptional control, we investigated the effects of deleting PCI8 on the total mRNA expression profile by oligonucleotide microarray analysis and found reduced mRNA levels for a subset of heat shock proteins in the pci8Delta mutant. We discuss possible dual functions of Pci8p and Int-6 in transcriptional and translational control.
Shalev
A
A
Laboratory of Gene Regulation and Development, NICHD, and the Basic Research Laboratory, NCI, National Institutes of Health, Bethesda, Maryland 20892, USA.
Valásek
L
L
Pise-Masison
C A
CA
Radonovich
M
M
Phan
L
L
Clayton
J
J
He
H
H
Brady
J N
JN
Hinnebusch
A G
AG
Asano
K
K
D001665:Binding Sites; D000075686:COP9 Signalosome Complex; D039621:Eukaryotic Initiation Factor-3 / Q000378:metabolism*; D005656:Fungal Proteins / Q000378:metabolism*; D020869:Gene Expression Profiling; D006801:Humans; D046912:Multiprotein Complexes; D010447:Peptide Hydrolases; D010448:Peptide Initiation Factors / Q000378:metabolism*; D039665:Prokaryotic Initiation Factor-3; D021122:Protein Subunits; D011506:Proteins / Q000502:physiology; D011518:Proto-Oncogene Proteins / Q000378:metabolism*; D012333:RNA, Messenger / Q000032:analysis; D012441:Saccharomyces cerevisiae / Q000737:chemistry*; D014158:Transcription, Genetic
C000589329:EIF3B protein, human; D039621:Eukaryotic Initiation Factor-3; D005656:Fungal Proteins; D046912:Multiprotein Complexes; D010448:Peptide Initiation Factors; D039665:Prokaryotic Initiation Factor-3; D021122:Protein Subunits; D011506:Proteins; D011518:Proto-Oncogene Proteins; D012333:RNA, Messenger; D010447:Peptide Hydrolases; D000075686:COP9 Signalosome Complex