Selective cleavage of BLM, the bloom syndrome protein, during apoptotic cell death.
Bloom syndrome (BS) is an autosomal recessive disorder characterized by a high incidence of cancer and genomic instability. BLM, the protein defective in BS, is a RECQ-like helicase that is presumed to function in mammalian DNA replication, recombination, or repair. We show here that BLM, but not the related RECQ-like helicase WRN, is rapidly cleaved in cells undergoing apoptosis. BLM was cleaved to 47- and 110-kDa major fragments, with kinetics similar to the apoptotic cleavage of poly(A)DP-ribose polymerase. BLM cleavage was prevented by a caspase 3 inhibitor and did not occur in caspase 3-deficient cells. Moreover, recombinant BLM was cleaved to 47- and 110-kDa fragments by caspase 3, but not caspase 6, in vitro. The caspase 3 recognition sequence (412)TEVD(415) was verified by mutating aspartate 415 to glycine and showing that this mutation rendered BLM resistant to caspase 3 cleavage. Cleavage did not abolish the BLM helicase activity but abolished BLM nuclear foci and the association of BLM with condensed DNA and the insoluble matrix. The results suggest that BLM, but not WRN, is an early selected target during the execution of apoptosis.
Bischof
O
O
Life Sciences Division, Lawrence Berkeley National Laboratory, Berkeley California 94720, USA.
Galande
S
S
Farzaneh
F
F
Kohwi-Shigematsu
T
T
Campisi
J
J
D000251:Adenosine Triphosphatases / Q000378:metabolism*; D017209:Apoptosis; D001483:Base Sequence; D053148:Caspase 3; D020169:Caspases / Q000378:metabolism; D004265:DNA Helicases / Q000378:metabolism*; D017931:DNA Primers; D005455:Fluorescent Antibody Technique; D006801:Humans; D006868:Hydrolysis; D019169:Jurkat Cells; D053484:RecQ Helicases
D017931:DNA Primers; C505407:CASP3 protein, human; D053148:Caspase 3; D020169:Caspases; D000251:Adenosine Triphosphatases; C097151:Bloom syndrome protein; D004265:DNA Helicases; D053484:RecQ Helicases