Abstract
Activation of c-Jun N-terminal kinase (JNK) by Fas ligation is caspase-dependent, suggesting that caspases may regulate activators of the JNK pathway. Here, we report that an upstream activator of JNK, hematopoietic progenitor kinase 1 (HPK1), was cleaved during apoptosis. Cleavage of HPK1 was blocked by peptide inhibitors for caspases. HPK1 was efficiently processed by recombinant caspase 3 in vitro. A conserved caspase recognition site, DDVD (amino acids 382 – 385), was found in the HPK1 protein sequence. By testing HPK1 proteins with in vivo and in vitro cleavage assays, we showed that aspartic acid residue 385 is the target for caspases. HPK1 cleavage separated the amino N-terminal kinase domain from the carboxyl C-terminal regulatory domain, and enhanced HPK1 kinase activity. Unlike the full-length HPK1, the N-terminal cleaved product failed to bind adaptor molecules Grb2 (growth factor receptor-bound protein 2) and Crk (CT10 regulator of kinase). The C-terminal fragment, although having three proline-rich domains, bound to Grb2 and Crk less efficiently than the full-length HPK1 protein. Taken together, the cleavage of HPK1 by caspase profoundly changed its biochemical properties.
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Abbreviations
- Crk:
-
CT10 regulator of kinase
- Grb2:
-
growth factor receptor bound protein 2
- GST:
-
glutathione S-transferase
- HPK1:
-
hematopoietic progenitor kinase 1
- JNK:
-
c-Jun N-terminal kinase
- kD:
-
kilodalton
- MAPK:
-
mitogen-activated protein kinase
- MBP:
-
myelin basic protein
- MEKK1:
-
MAPK kinase kinase 1
- MKK:
-
MAPK kinase
- PAK:
-
p21-GTPase-activated kinases
- z-DEVD-FK:
-
z-Asp-Glu-Val-Asp-fluoromethyl ketone
- z-VAD-FK:
-
z-Val-Ala-Asp-FK
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Acknowledgements
We thank Drs SM Feller, MC-T Hu, and EY Skolnik for generous gifts, members of Tan laboratory for their helpful discussions and critical reading of this manuscript, S Lee and R Afshar for technical assistance, and M Lowe for secretarial assistance. This work was supported by the National Institutes of Health grants R01-AI38649 and R01-AI42532 (to T-H Tan). T-H Tan is a Scholar of the Leukemia Society of America. Y-R Chen was supported by a Department of Defense Predoctoral Fellowship (DAMD17-97-1-7078) in the Breast Cancer Research Program, and is a recipient of Department of Defense Postdoctoral Fellowship (DAMD17-99-1-9507) in the Prostate Cancer Research Program. CF Meyer was supported by an NIH postdoctoral Trainingship in Immunology.
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Chen, YR., Meyer, C., Ahmed, B. et al. Caspase-mediated cleavage and functional changes of hematopoietic progenitor kinase 1 (HPK1). Oncogene 18, 7370–7377 (1999). https://doi.org/10.1038/sj.onc.1203116
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DOI: https://doi.org/10.1038/sj.onc.1203116
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