Subcellular localization of protein kinase C delta and epsilon affects transcriptional and post-transcriptional processes in four-cell mouse embryos.
During mouse preimplantation development, two isozymes of protein kinase C (PKC), delta and epsilon, transiently localize to nuclei at the early four-cell stage. In order to study their functions at this stage, we altered the subcellular localization of these isozymes (ratio of nuclear to cytoplasmic concentrations) with peptides that specifically activate or inhibit translocation of each isozyme. The effects of altering nuclear concentration of each isozyme on transcription (5-bromouridine 5'-triphosphate (BrUTP) incorporation), amount and distribution of small nuclear ribonucleoproteins (snRNPs), nucleolar dynamics (immunocytochemistry for Smith antigen (Sm) protein) and the activity of embryonic alkaline phosphatase (EAP; histochemistry) were examined. We found that nuclear concentration of PKC epsilon correlated with total mRNA transcription. Higher nuclear concentrations of both PKC delta and epsilon decreased storage of snRNPs in Cajal bodies and decreased the number of nucleoli, but did not affect the nucleoplasmic concentration of snRNPs. Inhibiting translocation of PKC delta out of the nucleus at the early four-cell stage decreased cytoplasmic EAP activity, whereas inhibiting translocation of PKC epsilon increased EAP activity slightly. These results indicate that translocation of PKC delta and epsilon in and out of nuclei at the early four-cell stage in mice can affect transcription or message processing, and that sequestration of these PKC in nuclei can also affect the activity of a cytoplasmic protein (EAP).
Dehghani
Hesam
H
Department of Biomedical Sciences, University of Guelph, Guelph, Ontario, N1G 2W1 Canada.
Reith
Cara
C
Hahnel
Ann C
AC
D000469:Alkaline Phosphatase / Q000378:metabolism; D000818:Animals; D001324:Autoantigens; D001692:Biological Transport; D001757:Blastomeres / Q000201:enzymology*; D002466:Cell Nucleolus / Q000378:metabolism / Q000648:ultrastructure; D002467:Cell Nucleus / Q000201:enzymology; D002478:Cells, Cultured; D003593:Cytoplasm / Q000201:enzymology; D007150:Immunohistochemistry / Q000379:methods; D051379:Mice; D018613:Microscopy, Confocal; D051745:Protein Kinase C-delta / Q000032:analysis*; D051744:Protein Kinase C-epsilon / Q000032:analysis*; D017411:Ribonucleoproteins, Small Nuclear / Q000032:analysis / Q000378:metabolism; D017381:Spliceosomes / Q000378:metabolism; D014158:Transcription, Genetic; D014178:Translocation, Genetic; D055517:snRNP Core Proteins
D001324:Autoantigens; D017411:Ribonucleoproteins, Small Nuclear; D055517:snRNP Core Proteins; D051745:Protein Kinase C-delta; D051744:Protein Kinase C-epsilon; D000469:Alkaline Phosphatase