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S‐adenosylmethionine and its metabolite induce apoptosis in HepG2 cells: Role of protein phosphatase 1 and Bcl‐x S
Author(s) -
Yang Heping,
Sadda Mamatha R.,
Li Mei,
Zeng Ying,
Chen Lixin,
Bae Wanjun,
Ou Xiaopeng,
Runnegar Maria T.,
Mato José M.,
Lu Shelly C.
Publication year - 2004
Publication title -
hepatology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 5.488
H-Index - 361
eISSN - 1527-3350
pISSN - 0270-9139
DOI - 10.1002/hep.20274
Subject(s) - protein phosphatase 1 , phosphatase , biology , apoptosis , dephosphorylation , microbiology and biotechnology , okadaic acid , messenger rna , phosphorylation , biochemistry , gene
S-adenosylmethionine (SAMe) and its metabolite 5'-methylthioadenosine (MTA) are proapoptotic in HepG2 cells. In microarray studies, we found SAMe treatment induced Bcl-x expression. Bcl-x is alternatively spliced to produce two distinct mRNAs and proteins, Bcl-x(L) and Bcl-x(S). Bcl-x(L) is antiapoptotic, while Bcl-x(S) is proapoptotic. In this study we showed that SAMe and MTA selectively induced Bcl-x(S) in a time- and dose-dependent manner in HepG2 cells. There are three transcription start sites in the human Bcl-x gene which yield only Bcl-x(L) in control HepG2 cells. SAMe and MTA treatment did not affect promoter usage, but while one promoter yielded only Bcl-x(L), the other two yielded both Bcl-x(L) and Bcl-x(S), with Bcl-x(S) as the predominant messenger RNA (mRNA) species. Trichostatin A, 3-deaza-adenosine, cycloleucine, and okadaic acid had no effect on Bcl-x(S) induction by SAMe or MTA. Calyculin A and tautomycin, on the other hand, blocked SAMe and MTA-mediated Bcl-x(S) induction and apoptosis in a dose-dependent manner. SAMe and MTA increased protein phosphatase 1 (PP1) catalytic subunit mRNA and protein levels and dephosphorylation of serine-arginine proteins, with the latter blocked by calyculin A. The effects of SAMe and MTA on Bcl-x(S), PP1 expression, and apoptosis were also seen in 293 cells, but not in primary hepatocytes. Induction of Bcl-x(S) by ceramide in HepG2 cells also resulted in apoptosis. In conclusion, we have uncovered a highly novel action of SAMe and MTA, namely the ability to affect the cellular phosphorylation state and alternative splicing of genes, in this case resulting in the induction of Bcl-x(S) leading to apoptosis.