Activation of mammalian ribosomal gene transcription requires phosphorylation of the nucleolar transcription factor UBF

Nucleic Acids Res. 1995 Jul 25;23(14):2593-9. doi: 10.1093/nar/23.14.2593.

Abstract

The nucleolar factor UBF is phosphorylated by casein kinase II (CKII) at serine residues within the C-terminal acidic domain which is required for transcription activation. To investigate the biological significance of UBF modification, we have compared the trans-activating properties of cellular UBF and recombinant UBF expressed in Escherichia coli. Using a variety of assays we demonstrate that unphosphorylated UBF is transcriptionally inactive and has to be phosphorylated at multiple sites to stimulate transcription. Examination of cDNA mutants in which the serine residues within the C-terminal domain were altered by site-directed mutagenesis demonstrates that CKII-mediated phosphorylations of UBF contribute to, but are not sufficient for, transcriptional activation. Besides CKII, other cellular protein kinases phosphorylate UBF at distinct sites in a growth-dependent manner. The marked differences in the tryptic peptide maps of UBF from growing and serum-starved cells suggest that alterations in the degree of UBF phosphorylation may modulate rRNA synthetic activity in response to extracellular signals.

Publication types

  • Comparative Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • 3T3 Cells
  • Amino Acid Sequence
  • Animals
  • Base Sequence
  • Casein Kinase II
  • Cell Division
  • Cell Nucleolus / metabolism
  • DNA Primers / genetics
  • DNA, Complementary / genetics
  • DNA, Ribosomal / genetics*
  • DNA-Binding Proteins / genetics
  • DNA-Binding Proteins / metabolism*
  • Mice
  • Molecular Sequence Data
  • Mutagenesis, Site-Directed
  • Peptide Mapping
  • Phosphorylation
  • Pol1 Transcription Initiation Complex Proteins*
  • Protein-Serine-Threonine Kinases / metabolism
  • Recombinant Proteins / genetics
  • Recombinant Proteins / metabolism
  • Transcription Factors / genetics
  • Transcription Factors / metabolism*
  • Transcriptional Activation

Substances

  • DNA Primers
  • DNA, Complementary
  • DNA, Ribosomal
  • DNA-Binding Proteins
  • Pol1 Transcription Initiation Complex Proteins
  • Recombinant Proteins
  • Transcription Factors
  • transcription factor UBF
  • Casein Kinase II
  • Protein-Serine-Threonine Kinases