Nucleic Acids Research, 2000, Vol. 28, No. 21 4180-4188
© 2000 Oxford University Press
Proteolysis of the human DNA polymerase 
catalytic subunit by caspase-3 and calpain specifically during apoptosis
Division of Biochemistry and Molecular Biology, 229 Stanley Hall, University of California, Berkeley, CA 94720-3206, USA
Human DNA polymerase epsilon (pol 
) normally contains a 261-kDa catalytic subunit (p261), but from some sources it is isolated as a 140-kDa catalytic core of p261. This shortened form possesses normal or somewhat enhanced polymerase activity and its significance is unknown. We report here that caspase-3 and calpain can form p140 from p261 in vitro and in vivo and that during early stages of apoptosis induced in Jurkat cells by staurosporine or anti-Fas-activating antibody, p261 is cleaved into p140 by caspase-3. At later stages, activated calpain might also contribute to this conversion. The sites of cleavage by caspase-3 have been identified, and mutations at these ‘DEAD boxes’ resulted in cleavage-resistant enzyme. Cleavage at these sites separates the ‘N-terminal catalytic core’ from the ‘C-terminal’ regions described for p261. Cleavage does not occur during necrosis or following exposure to H2O2 or methanesulfonic acid methyl ester. p140 is unlikely to be able to functionally replace p261 in vivo, since it does not bind to PCNA or the other pol subunits.
* To whom correspondence should be addressed. Tel: +1 510 642 7583; Fax: +1 510 643 9290; Email: slinn@socrates.berkeley.edu Present address: Wei Liu, Genetics Institute, MS 35–1127, 35 Cambridgepark, MA 02140, USA
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