O-GlcNAc cycling enzymes associate with the translational machinery and modify core ribosomal proteins

Quira Zeidan, Zihao Wang, Antonio De Maio, Gerald Warren Hart

Research output: Contribution to journalArticle

Abstract

Protein synthesis is globally regulated through posttranslational modifications of initiation and elongation factors. Recent high-throughput studies have identified translation factors and ribosomal proteins (RPs) as substrates for the O-GlcNAc modification. Here we determine the extent and abundance of O-GlcNAcylated proteins in translational preparations. O-GlcNAc is present on many proteins that form active polysomes. We identify twenty O-GlcNAcylated core RPs, of which eight are newly reported. We map sites of O-GlcNAc modification on four RPs (L6, L29, L32, and L36). RPS6, a component of the mammalian target of rapamycin (mTOR) signaling pathway, follows different dynamics of OGlcNAcylation than nutrient-induced phosphorylation. We also show that both O-GlcNAc cycling enzymes OGT and OGAse strongly associate with cytosolic ribosomes. Immunofluorescence experiments demonstrate that OGAse is present uniformly throughout the nucleus, whereas OGT is excluded from the nucleolus. Moreover, nucleolar stress only alters OGAse nuclear staining, but not OGT staining. Lastly, adenovirus-mediated overexpression of OGT, but not of OGAse or GFP control, causes an accumulation of 60S subunits and 80S monosomes. Our results not only establish that O-GlcNAcylation extensively modifies RPs, but also suggest that O-GlcNAc play important roles in regulating translation and ribosome biogenesis.

Original languageEnglish (US)
Pages (from-to)1922-1936
Number of pages15
JournalMolecular Biology of the Cell
Volume21
Issue number12
DOIs
StatePublished - Jun 15 2010

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Ribosomal Proteins
Ribosomes
Enzymes
Staining and Labeling
Peptide Elongation Factors
Peptide Initiation Factors
Proteins
Polyribosomes
Sirolimus
Post Translational Protein Processing
Adenoviridae
Fluorescent Antibody Technique
Phosphorylation
Food

ASJC Scopus subject areas

  • Molecular Biology
  • Cell Biology
  • Medicine(all)

Cite this

O-GlcNAc cycling enzymes associate with the translational machinery and modify core ribosomal proteins. / Zeidan, Quira; Wang, Zihao; De Maio, Antonio; Hart, Gerald Warren.

In: Molecular Biology of the Cell, Vol. 21, No. 12, 15.06.2010, p. 1922-1936.

Research output: Contribution to journalArticle

Zeidan, Quira ; Wang, Zihao ; De Maio, Antonio ; Hart, Gerald Warren. / O-GlcNAc cycling enzymes associate with the translational machinery and modify core ribosomal proteins. In: Molecular Biology of the Cell. 2010 ; Vol. 21, No. 12. pp. 1922-1936.
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