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 b y the cell shape-modulating fungal metabolite cytochalasin D (CD). Ind uction 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 clo sely correlated with increases in 43 kDa p52(PAI-1) core protein biosy nthesis. Sensitivity to tunicamycin indicated that N-linked post-trans lational modifications to this 43 kDa core species generated the full complement of 50 kDa (intermediate) and 52 kDa (mature) p52(PAI-1) gly cosylated isoforms. CD-induced p52(PAI-1) expression occurred efficien tly in quiescent NRK cells maintained under serum-free conditions as w ell as in fully serum-supplemented actively growing cultures. While 8- bromo-cAMP reduced both constitutive and transforming growth factor-be ta-induced p52(PAI-1) synthesis by >50%, no such inhibition was eviden t in short-term (4 h) CD-stimulated cultures. Long-term (24 h) exposur e of NRK/CD cells to 8-bromo-cAMP did result in an approximately 34% r eduction in stimulated p52(PAI-1) expression, however, levels expresse d by NRK/CD + cAMP populations remained markedly elevated relative to control values. These data suggest the existence of a cell shape-depen dent aspect of p52(PAI-1) expression control distinct from both the co nstitutive and growth factor-mediated pathways of gene regulation. (C) 1994 Wiley-Liss, Inc.