Increases in mRNA concentrations of the alpha and beta subunits of prolyl 4-hydroxylase accompany increased gene expression of type IV collagen during differentiation of mouse F9 cells.
Mouse F9 teratocarcinoma stem cells differentiate in monolayer cultures in the presence of retinoic acid, dibutyryl cAMP, and isobutyl methylxanthine. This differentiation is associated with a marked increase in the synthesis rates and mRNA concentrations of basement membrane proteins such as type IV collagen. We report here that the differentiation also involves an increase of up to 50-fold in the concentrations of the mRNAs for the alpha and beta subunits of prolyl 4-hydroxylase, the enzyme required for the cotranslational and post-translational hydroxylation of proline residues in collagens. The time courses and magnitudes of increases in these two mRNA concentrations were similar to those observed in the same experiments for the mRNA of the alpha chain of type IV collagen. In the differentiated F9 cells the concentration of the alpha subunit mRNA was about 30% of the beta subunit mRNA concentration. Northern blot analyses indicated that the sizes of the alpha and beta subunit mRNAs in the differentiated mouse F9 cells are similar to those in human skin fibroblasts. The F9 cell differentiation system appears to provide a useful model for studies on the regulation of prolyl 4-hydroxylase synthesis.
Helaakoski
T
T
Department of Medical Biochemistry, University of Oulu, Finland.
Pajunen
L
L
Kivirikko
K I
KI
Pihlajaniemi
T
T
D015056:1-Methyl-3-isobutylxanthine / Q000494:pharmacology; D000818:Animals; D015152:Blotting, Northern; D003994:Bucladesine / Q000494:pharmacology; D002454:Cell Differentiation; D002460:Cell Line; D003094:Collagen / Q000235:genetics*; D015342:DNA Probes; D005786:Gene Expression Regulation; D005796:Genes; D005987:Glyceraldehyde-3-Phosphate Dehydrogenases / Q000235:genetics; D007700:Kinetics; D046911:Macromolecular Substances; D051379:Mice; D011393:Procollagen-Proline Dioxygenase / Q000235:genetics*; D012333:RNA, Messenger / Q000096:biosynthesis / Q000235:genetics* / Q000302:isolation & purification; D013724:Teratoma; D014212:Tretinoin / Q000494:pharmacology; D014407:Tumor Cells, Cultured / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism
D015342:DNA Probes; D046911:Macromolecular Substances; D012333:RNA, Messenger; D014212:Tretinoin; D003994:Bucladesine; D003094:Collagen; D011393:Procollagen-Proline Dioxygenase; D005987:Glyceraldehyde-3-Phosphate Dehydrogenases; D015056:1-Methyl-3-isobutylxanthine