Differential regulation of the stability of cyclic AMP-dependent protein kinase messenger RNA in normal versus neoplastic mouse lung epithelial cells.
Neoplastic mouse lung epithelial cells contain greatly diminished activity, protein, and mRNA for the type I isozyme of cyclic AMP-dependent protein kinase (PKA I), while expression of the type II isozyme (PKA II) is similar to that of normal lung cells. A time course of PKA mRNA content in transcriptionally inhibited cells indicated that most PKA mRNAs are more stable in the neoplastic E9 cell line than in related nontumorigenic C10 cells. To address the basis of this differential stability, we treated both cell lines with cycloheximide, an inhibitor of protein synthesis, in the presence or absence of the transcriptional inhibitor, 5,6-dichloro-1-b-ribofuranosyl-benzimidazole (DRB). The rate of PKA II regulatory subunit alpha mRNA decay in the presence of DRB was unaffected by cycloheximide treatment in E9 cells but decreased upon the addition of cycloheximide to DRB-treated C10 cells. The combination of these two agents markedly destabilized PKA II mRNAs (PKA catalytic subunit alpha and PKA II regulatory subunit alpha) relative to DRB treatment alone in neoplastic E9 cells, causing them to decay at a rate equal to that in C10 cells. PKA II mRNA may be specifically stabilized by a protein with a relatively short half-life in neoplastic E9 cells. These results suggest the involvement of tumor-specific factor(s) in the regulation of PKA mRNA stability, a potential mechanism for conferring the observed differential responsiveness of normal and neoplastic lung cells to cyclic AMP.
Lange-Carter
C A
CA
Molecular and Environmental Toxicology Program, Colorado Cancer Center, School of Pharmacy, University of Colorado, Boulder 80309-0297.
Malkinson
A M
AM
D000818:Animals; D015152:Blotting, Northern; D003513:Cycloheximide / Q000494:pharmacology; D004004:Dichlororibofuranosylbenzimidazole / Q000494:pharmacology; D004848:Epithelium / Q000502:physiology; D015870:Gene Expression / Q000187:drug effects; D008168:Lung / Q000502:physiology*; D008175:Lung Neoplasms / Q000235:genetics*; D051379:Mice; D008807:Mice, Inbred BALB C; D011494:Protein Kinases / Q000235:genetics*; D012333:RNA, Messenger / Q000378:metabolism*; D012334:RNA, Neoplasm / Q000378:metabolism*
D012333:RNA, Messenger; D012334:RNA, Neoplasm; D004004:Dichlororibofuranosylbenzimidazole; D003513:Cycloheximide; D011494:Protein Kinases