Cell cycle regulation of the glyceraldehyde-3-phosphate dehydrogenase/uracil DNA glycosylase gene in normal human cells

Nucleic Acids Res. 1993 Feb 25;21(4):993-8. doi: 10.1093/nar/21.4.993.

Abstract

The cell cycle regulation of the glyceraldehyde-3-phosphate dehydrogenase (GAPDH)/uracil DNA glycosylase (UDG) gene was examined in normal human cells. Steady state RNA levels were monitored by Northern blot analysis using a plasmid (pChug 20.1) which contained the 1.3 kb GAPDH/UDG cDNA. The biosynthesis of the 37 kDa GAPDH/UDG protein was determined using an anti-human placental GAPDH/UDG monoclonal antibody to immunoprecipitate the radiolabeled protein. Increases in steady state GAPDH/UDG mRNA levels were cell cycle specific. A biphasic pattern was observed resulting in a 19-fold increase in the amount of GAPDH/UDG mRNA. The biosynthesis of the 37 kDa GAPDH/UDG protein displayed a similar biphasic regulation with a 7-fold increase. Pulse-chase experiments revealed a remarkably short half life of less than 1 hr. for the newly synthesized 37 kDa protein, comparable to that previously documented for a number of oncogenes. GAPDH/UDG mRNA levels were markedly reduced at 24 hr. when DNA synthesis was maximal. These results define the GAPDH/UDG gene as cell cycle regulated with a characteristic temporal sequence of expression in relation to DNA synthesis. The cell cycle synthesis of a labile 37 kDa monomer suggests a possible regulatory function for this multidimensional protein. Further, modulation of the GAPDH/UDG gene in the cell cycle may preclude its use as a reporter gene when the proliferative state of the cell is not kept constant.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Cell Cycle / genetics*
  • Cells, Cultured
  • DNA Glycosylases*
  • Gene Expression Regulation, Enzymologic
  • Glyceraldehyde-3-Phosphate Dehydrogenases / biosynthesis
  • Glyceraldehyde-3-Phosphate Dehydrogenases / genetics*
  • Humans
  • Molecular Sequence Data
  • Multienzyme Complexes / biosynthesis
  • Multienzyme Complexes / genetics
  • N-Glycosyl Hydrolases / biosynthesis
  • N-Glycosyl Hydrolases / genetics*
  • RNA, Messenger / metabolism
  • S Phase
  • Uracil-DNA Glycosidase

Substances

  • Multienzyme Complexes
  • RNA, Messenger
  • Glyceraldehyde-3-Phosphate Dehydrogenases
  • DNA Glycosylases
  • N-Glycosyl Hydrolases
  • Uracil-DNA Glycosidase

Associated data

  • GENBANK/X53778