Induced expression of p52(PAI-1) in normal rat kidney cells by the microfilament-disrupting agent cytochalasin D.
In established normal rat kidney (NRK) cells, synthesis of the 52 kDa type-1 inhibitor of plasminogen activator [p52(PAI-1)] is stimulated by the cell shape-modulating fungal metabolite cytochalasin D (CD). Induction paralleled the time course of morphologic change and reflected relatively specific increases in saponin-resistant p52(PAI-1) protein accumulation (approximating ten- to thirty-fold over control) and mRNA abundance (seven- to nine-fold). Augmented p52 (PAI-1) mRNA levels closely correlated with increases in 43 kDa p52(PAI-1) core protein biosynthesis. Sensitivity to tunicamycin indicated that N-linked post-translational modifications to this 43 kDa core species generated the full complement of 50 kDa (intermediate) and 52 kDa (mature) p52(PAI-1) glycosylated isoforms. CD-induced p52(PAI-1) expression occurred efficiently in quiescent NRK cells maintained under serum-free conditions as well as in fully serum-supplemented actively growing cultures. While 8-bromo-cAMP reduced both constitutive and transforming growth factor-beta-induced p52(PAI-1) synthesis by > 50%, no such inhibition was evident in short-term (4 h) CD-stimulated cultures. Long-term (24 h) exposure of NRK/CD cells to 8-bromo-cAMP did result in an approximately 34% reduction in stimulated p52(PAI-1) expression, however, levels expressed by NRK/CD+cAMP populations remained markedly elevated relative to control values. These data suggest the existence of a cell shape-dependent aspect of p52(PAI-1) expression control distinct from both the constitutive and growth factor-mediated pathways of gene regulation.
Higgins
P J
PJ
Department of Microbiology, Immunology, and Molecular Genetics, Albany Medical College, New York 12208.
Ryan
M P
MP
Providence
K M
KM
D015124:8-Bromo Cyclic Adenosine Monophosphate / Q000494:pharmacology; D000818:Animals; D015152:Blotting, Northern; D002460:Cell Line; D015638:Cytochalasin D / Q000494:pharmacology*; D004591:Electrophoresis, Polyacrylamide Gel; D015870:Gene Expression / Q000235:genetics; D007668:Kidney / Q000737:chemistry / Q000166:cytology*; D017395:Plasminogen Activator Inhibitor 1 / Q000032:analysis / Q000235:genetics / Q000502:physiology*; D012333:RNA, Messenger / Q000032:analysis / Q000235:genetics; D051381:Rats; D013997:Time Factors; D014415:Tunicamycin / Q000494:pharmacology
D017395:Plasminogen Activator Inhibitor 1; D012333:RNA, Messenger; D014415:Tunicamycin; D015638:Cytochalasin D; D015124:8-Bromo Cyclic Adenosine Monophosphate