PKR Regulates B56α-mediated BCL2 Phosphatase Activity in Acute Lymphoblastic Leukemia-derived REH Cells

• Published in: The Journal of biological chemistry 2008-12, Vol.283 (51), p.35474-35485
• Main Authors: Ruvolo, Vivian R., Kurinna, Svitlana M., Karanjeet, Kul B., Schuster, Todd F., Martelli, Alberto M., McCubrey, James A., Ruvolo, Peter P.
• Format: Article
• Language: English
• Subjects: Mechanisms of Signal Transduction
• ISSN: 0021-9258; 1083-351X
• DOI: 10.1074/jbc.M800951200

Protein phosphatase 2A (PP2A) is a heterotrimer comprising catalytic, scaffold, and regulatory (B) subunits. There are at least 21 B subunit family members. Thus PP2A is actually a family of enzymes defined by which B subunit is used. The B56 family member B56α is a phosphoprotein that regulates dephosphorylation of BCL2. The stress kinase PKR has been shown to phosphorylate B56α at serine 28 in vitro, but it has been unclear how PKR might regulate the BCL2 phosphatase. In the present study, PKR regulation of B56α in REH cells was examined, because these cells exhibit robust BCL2 phosphatase activity. PKR was found to be basally active in REH cells as would be predicted if the kinase supports B56α-mediated dephosphorylation of BCL2. Suppression of PKR promoted BCL2 phosphorylation with concomitant loss of B56α phosphorylation at serine 28 and inhibition of mitochondrial PP2A activity. PKR supports stress signaling in REH cells, as suppression of PKR promoted chemoresistance to etoposide. Suppression of PKR promoted B56α proteolysis, which could be blocked by a proteasome inhibitor. However, the mechanism by which PKR supports B56α protein does not involve PKR-mediated phosphorylation of the B subunit at serine 28 but may involve eIF2α activation of AKT. Phosphorylation of serine 28 by PKR promotes mitochondrial localization of B56α, because wild-type but not mutant S28A B56α promoted mitochondrial PP2A activity. Cells expressing wild-type B56α but not S28A B56α were sensitized to etoposide. These results suggest that PKR regulates B56α-mediated PP2A signaling in REH cells.