Induction of murine teratocarcinoma cell differentiation by suppression of poly(ADP-ribose) synthesis.

Abstract

Poly(ADP-ribose) synthesizing activity in mouse teratocarcinoma EC-Al cells decreased markedly during differentiation induced by retinoic acid; the activities assayed in permeabilized cells decreased to 25% and 10% of the activity of control (uninduced cells) 2 and 3 days, respectively, after the addition of 0.1 ,uM retinoic acid to the culture medium. This change preceded changes in morphology and DNA synthesis, which became prominent after 4 days. The decrease in poly(ADP-ribose) synthesizing activity appeared to be caused by a diminution of the synthetase protein and not by a decrease in its catalytic activity, because the full activity disclosed by DNase I treatment decreased in parallel, albeit at about 20 times higher levels. When 8 mM 3-aminobenzamide or 10 mM nicotinamide, specific inhibitors of poly(ADP-ribose) synthetase, was added to the culture medium, the cells underwent differentiation after 7-9 days. An analogue, 3-aminobenzoic acid, which is not inhibitory to the synthetase, induced differentiation much less efficiently than did 3-aminobenzamide, and the effect of 3-aminobenzoic acid appeared to be ascribable to its potent cytotoxicity. Immunohistochemical analysis using anti-poly(ADP-ribose) antibody confirmed the marked reduction in poly(ADP-ribose) synthesizing activity in nuclei of the cells treated with retinoic acid or 3-aminobenzamide but not with 3-aminobenzoic acid. These results suggest that a decrease in poly(ADP-ribose) synthesis triggers differentiation of teratocarcinoma cells.Retinoic acid and related compounds (retinoids) have been shown to induce differentiation of several lines of murine teratocarcinoma cells into endoderm epitheloid cells or fibroblast-like cells (1, 2). The molecular mechanism of this induction of differentiation is not yet fully understood (3); De Luca (4), Wolf et al. (5), and Jetten and De Luca (6) reported observations suggesting a role of retinyl phosphate mannose in early cell surface changes, while Chytil and Ong (7) and Jetten and Jetten (8) proposed an alteration in transcription mediated by specific binding proteins. We became interested in these cells in view of our and others' finding that poly(ADP-ribose) is closely related to cell differentiation (9, 10). Poly(ADP-ribose) is a macromolecule synthesized from NAD in eukaryotic cells by a chromatin-bound enzyme, poly(ADP-ribose) synthetase (11). This synthesis proceeds in covalent attachment, at one terminus of the polymer, to various protein acceptors such as histones, nonhistone chromosomal proteins, and the synthetase itself (11). Accumulating evidence suggests that this polymer is involved in DNA repair (12-14), cell transformation (15, 16), and cell differentiation (9,10). Previously, we demonstrated that the poly(ADP-ribose) synthesizing activity serves as a marker of differentiation of human granulocytes (17, 18), leukemic lymphocytes (19), and epidermal cells (20

Nicotinamide is a potent inducer of endocrine differentiation in cultured human fetal pancreatic cells.

“…The activity ofpoly(ADP-ribose) synthetase is linked with the level of cellular differentiation in several cell types. Evidence for this comes from experiments with preadipocytes (41), intestinal epithelial cells (42), chondrocytes (43), teratocarcinoma cells (44), and thymocytes (45). In all of these cells, the ADP-ribosylating activity is lower in the more differentiated phenotype, and morphological evidence of differentiation is preceded by diminished activity of poly(ADP-ribose) synthetase.…”

“…In all of these cells, the ADP-ribosylating activity is lower in the more differentiated phenotype, and morphological evidence of differentiation is preceded by diminished activity of poly(ADP-ribose) synthetase. Furthermore, chondrocyte and teratocarcinoma cell differentiation can be triggered by inhibitors of the enzyme, such as NIC and 3-aminobenzamide (44,46). These effects are likely to reflect changes in the ADP ribosylation of nuclear proteins, resulting in modifications in chromatin structure that enable the transcription of previously masked regions of DNA.…”

Functional interaction between human papillomavirus type 18 E2 and poly(ADP-ribose) polymerase 1

“…Further studies for the regulation of host gene expression by E2 is required for understanding E2-mediated replicative senescence. It is known that PARP-1 is able to modulate cell differentiation and tumorigenesis (Ohashi et al, 1984;Masutani et al, 2001;Smulson et al, 1995). HPV oncoproteins can modulate the ability of primary human keratinocytes to differentiate (Funk et al, 1997;Jones et al, 1999;Sherman and Schlegel, 1996;Sherman et al, 1997).…”

ADP‐Ribosylation of Histones in Nuclei Isolated from Embryos of the Sea Urchin, Hemicentrotus pulcherrimus

“…ADP-ribosylation of these nuclear proteins is also thought to play important roles in cell proliferation and differentiation (11,14,17,24). In sea urchin embryos, a marked increase followed by a sudden decrease in the rate of ADP-ribosylation occurs in the pre-hatching period and also in the post-hatching period, with peaks of the rate at the morula and gastrula stages (10).…”