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Eukaryotic rRNA Modification by Yeast 5-Methylcytosine-Methyltransferases and Human Proliferation-Associated Antigen

Gabrielle Bourgeois1, Michel Ney1, Imre Gaspar2

  • 1Laboratoire IMoPA, UMR 7365 UL-CNRS, BioPole de UL, Vandoeuvre-les-Nancy, France.

Plos One
|July 22, 2015
PubMed

Insights

Human p120 protein exhibits RNA:m5C-methyltransferase activity, restoring 5-methylcytosine (m5C) in yeast rRNA. This enzyme plays a role in pre-ribosomal RNA processing and quality control during ribosome synthesis.

Area of Science:

  • Molecular Biology
  • Biochemistry
  • RNA Biology

Background:

  • 5-methylcytosine (m5C) is a crucial RNA modification found in various eukaryotic RNAs.
  • RNA:m5C-methyltransferases (MTases), including yeast Nop2 and human p120, are key enzymes in this process.
  • Human p120 is recognized as a tumor marker overexpressed in cancers.

Purpose of the Study:

  • To investigate the RNA:m5C-MTase activity of human p120.
  • To elucidate the roles of Nop2 and p120 in rRNA modification and pre-ribosomal RNA processing.
  • To understand the functional importance of protein domains in Nop2 and p120.

Main Methods:

  • RNA bisulfite sequencing
  • High-Performance Liquid Chromatography with tandem Mass Spectrometry (HPLC-MS/MS)
  • Functional complementation assays in yeast
  • Chimeric protein analysis

Main Results:

  • Human p120 demonstrated RNA:m5C-MTase activity, restoring m5C at position 2870 in 25S rRNA.
  • Yeast Nop2p and Rcm1p catalyze m5C formation in specific rRNA domains; no m5C found in 18S rRNA.
  • Nop2 N-terminal domain is critical for protein localization and function; Nop2, not m5C, is essential for pre-rRNA processing.

Conclusions:

  • Human p120 possesses RNA:m5C-MTase activity, functionally complementing yeast Nop2.
  • Nop2 functions as a pre-ribosomal protein involved in both rRNA processing and m5C modification.
  • These findings highlight the role of m5C modification enzymes in ribosome biogenesis quality control.

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