Protein kinase NII and the regulation of rDNA transcription in mammalian cells.
Transcription of ribosomal RNA genes is generally accepted to correlate with cell growth. Using primary cultures of adult bovine aortic endothelial (ABAE) cells, we have shown that transcription of rDNA in confluent cells falls to 5% of the transcription level in growing cells. Protein kinase NII appears to be a limiting factor to promote rDNA transcription in isolated nuclei of confluent cells. Protein kinase NII was detected by immunocytochemistry in the cytoplasm, nuclei and nucleoli of growing cells while it was no longer present in nucleoli of confluent cells. The kinase activity, in isolated nuclei, was estimated by endogenous phosphorylation of a specific substrate, nucleolin. A 10% residual activity was present in confluent cell nuclei compared to growing cell nuclei. Concomitantly, the transcription 'in vitro' of rDNA in the corresponding nuclei was also highly reduced (by 85%). Addition of exogenous protein kinase NII to confluent cell nuclei induced a strong increase in the phosphorylation of specific proteins including nucleolin. In parallel, the transcription of rDNA was increased by a factor of 5, to nearly the level observed in nuclei prepared from growing cells. These data suggest that, in confluent cells, factors necessary for rDNA transcription machinery are present but inactive in the nucleolus and that the phosphorylation of one or several of these factors (nucleolin, topoisomerase I,...) by protein kinase NII is a key event in the regulation of rDNA transcription.
Centre de Recherche de Biochimie et de Génétique Cellulaires du CNRS, Toulouse, France.
P
P
Belenguer
V
V
Baldin
C
C
Mathieu
H
H
Prats
M
M
Bensaid
G
G
Bouche
F
F
Amalric
D000469:Alkaline Phosphatase / Q000378:metabolism; D000818:Animals; D002417:Cattle; D002455:Cell Division; D002478:Cells, Cultured; D004275:DNA, Ribosomal / Q000235:genetics*; D004730:Endothelium, Vascular / Q000166:cytology / Q000201:enzymology / Q000378:metabolism*; D005809:Genes, Regulator; D007700:Kinetics; D009687:Nuclear Proteins / Q000378:metabolism*; D010750:Phosphoproteins / Q000378:metabolism; D010766:Phosphorylation; D011494:Protein Kinases / Q000378:metabolism*; D016601:RNA-Binding Proteins; D014158:Transcription, Genetic; D000097796:Nucleolin
D000469:Alkaline Phosphatase; D004275:DNA, Ribosomal; D009687:Nuclear Proteins; D010750:Phosphoproteins; D011494:Protein Kinases; D016601:RNA-Binding Proteins; D000097796:Nucleolin; C026813:protein kinase NII