Inhibitory effect of calcium-binding protein regucalcin on ribonucleic acid synthesis in isolated rat liver nuclei.

The effect of regucalcin, a Ca2+-binding protein isolated from rat liver cytosol, on ribonucleic acid (RNA) synthesis in the nuclei of normal rat liver and of regenerating rat liver was investigated. The liver weight at 1 day after partial hepatectomy was increased about 50% of that of sham-operated (control) rats. Calcium chloride (1.0-20 microM Ca2+ as final concentration) was added into the reaction mixture of nuclear RNA synthesis. RNA synthesis was established by incorporation of [3H]-uridine 5'-triphosphate (UTP) into the nuclear RNA. Addition of Ca2+ (5 and 10 microM) caused a significant increase of RNA synthesis in the nuclei from control rat liver. Such effect of Ca2+ was potentiated in the nuclei of regenerating liver; nuclear RNA synthesis was increased about 2 fold by the 1.0 and 2.5 microM Ca2+ addition. The stimulatory effect of Ca2+ was significantly inhibited by the presence of alpha-amanitin (10(-8) M), an inhibitor of RNA polymerase II. The presence of regucalcin (0.25 and 0.5 microM) significantly inhibited RNA synthesis in the nuclei from control rat liver and from regenerating rat liver. The inhibitory effect of regucalcin was remarkable in the presence of EGTA (0.5 mM), and it was weakened by the addition of Ca2+ (5 microM). Such regucalcin effect was not seen in the presence of alpha-amanitin. The presence of anti-regucalcin IgG in the reaction mixture significantly increased RNA synthesis in the nuclei from control rat liver, indicating that the endogenous regucalcin may be involved in nuclear RNA synthesis. The present results demonstrate that regucalcin can inhibit nuclear RNA synthesis in rat liver. Regucalcin may have an inhibitory role in the regulation of liver nuclear RNA synthesis.