Int6 regulates both proteasomal degradation and translation initiation and is critical for proper formation of acini by human mammary epithelium

J. Suo, S. J. Snider, Gordon Mills, C. J. Creighton, A. C. Chen, R. Schiff, R. E. Lloyd, E. C. Chang

Research output: Contribution to journalArticle

16 Citations (Scopus)

Abstract

INT6/EIF3E has been implicated in breast tumorigenesis, but its functional activities remain poorly defined. We found that, repressing INT6 expression induced transformed properties in normal human mammary epithelium (MCF10A); in contrast, Int6 silencing induced apoptosis in HeLa cells. As in fission yeast, Int6 in human cells was required for assembly of active proteasomes. A reverse-phase protein array screen identified SRC3/AIB1 as one oncoprotein the level and stability of which increased when Int6 was silenced in MCF10A cells. Our data further show that Int6 binds SRC3 and its ubiquitin ligase Fbw7, thus perhaps mediating the interaction between SRC3-Fbw7 and proteasomes. Consistent with this, Int6 silencing did not increase SRC3 levels in HeLa cells, which have low Fbw7 levels. It is surprising that, however, polyubiquitylated proteins do not accumulate or may even decrease in Int6-silenced cells that contain defective proteasomes. Considering that decreased ubiquitin might explain this observation and that Int6 might control ubiquitin levels in its role as a subunit of eIF3 (eukaryote translation initiation factor 3), we found that silencing Int6 reduced monoubiquitin protein levels, which correlated with a shift of ubiquitin mRNAs from larger polysomes to non-translating ribosomes. In contrast, levels of many housekeeping proteins did not change. This apparent reduction in the translation of ubiquitin genes correlated with a modest reduction in protein synthesis rate and formation of large polysomes. To further determine whether Int6 can selectively control translation, we analyzed translation of different 5′-untranslated region reporters and found that indeed, loss of Int6 had differential effects on these reporters. Together the data suggest that Int6 depletion blocks ubiquitin-dependent proteolysis by decreasing both ubiquitin levels and the assembly of functional proteasome machinery, leading to accumulation of oncoproteins, such as SRC3 that can transform mammary epithelium. Our data also raise the possibility that Int6 can further fine-tune protein levels by selectively controlling translation of specific mRNAs.

Original languageEnglish (US)
Pages (from-to)724-736
Number of pages13
JournalOncogene
Volume30
Issue number6
DOIs
StatePublished - Feb 10 2011
Externally publishedYes

Fingerprint

Human Mammary Glands
Ubiquitin
Proteasome Endopeptidase Complex
Polyribosomes
Oncogene Proteins
Proteins
HeLa Cells
Prokaryotic Initiation Factor-3
Breast
Housekeeping
Protein Array Analysis
Schizosaccharomyces
5' Untranslated Regions
Protein Biosynthesis
Ligases
Eukaryota
Ribosomes
Proteolysis
Carcinogenesis
Epithelium

Keywords

  • eIF3
  • proteasomes
  • translation
  • tumor suppressor
  • ubiquitin

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics
  • Cancer Research

Cite this

Int6 regulates both proteasomal degradation and translation initiation and is critical for proper formation of acini by human mammary epithelium. / Suo, J.; Snider, S. J.; Mills, Gordon; Creighton, C. J.; Chen, A. C.; Schiff, R.; Lloyd, R. E.; Chang, E. C.

In: Oncogene, Vol. 30, No. 6, 10.02.2011, p. 724-736.

Research output: Contribution to journalArticle

Suo, J. ; Snider, S. J. ; Mills, Gordon ; Creighton, C. J. ; Chen, A. C. ; Schiff, R. ; Lloyd, R. E. ; Chang, E. C. / Int6 regulates both proteasomal degradation and translation initiation and is critical for proper formation of acini by human mammary epithelium. In: Oncogene. 2011 ; Vol. 30, No. 6. pp. 724-736.
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AU - Chen, A. C.

AU - Schiff, R.

AU - Lloyd, R. E.

AU - Chang, E. C.

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