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  • Original Paper
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Caspase-mediated cleavage and functional changes of hematopoietic progenitor kinase 1 (HPK1)

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

References

  • Anafi M, Kiefer F, Gish GD, Mbamalu G, Iscove NN and Pawson T. . 1997 J. Biol. Chem. 272: 27804–27811.

  • Bagrodia S, Derijard B, Davis RJ and Ceroine RA. . 1995 J. Biol. Chem. 270: 27995–27998.

  • Brenner B, Koppenhoefer U, Weinstock C, Linderkamp O, Lang F and Gulbins E. . 1997 J. Biol. Chem. 272: 22173–22181.

  • Cardone M, Salvesen GS, Widmann C, Johnson G and Frisch SM. . 1997 Cell 90: 315–323.

  • Chen Y-R, Meyer CF and Tan T-H. . 1996a J. Biol. Chem. 271: 631–634.

  • Chen Y-R, Wang W, Kong A-NT and Tan T-H. . 1998 J. Biol. Chem. 273: 1769–1775.

  • Chen Y-R, Wang X, Templeton D, Davis RJ and Tan T-H. . 1996b J. Biol. Chem. 271: 31929–31936.

  • Deak JC, Cross JV, Lewis M, Qian Y, Parrott LA, Distelhorst CW and Templeton DJ. . 1998 Proc. Natl. Acad. Sci. USA 95: 5595–5600.

  • Diener K, Wang XS, Chen C, Meyer CF, Bray J, Zukowski M, Tan T-H and Yao Z. . 1997 Proc. Natl. Acad. Sci. USA 94: 9687–9692.

  • Emoto Y, Manome Y, Meinhardt G, Kisaki H, Kharbabda S, Robertson M, Ghayur T, Wong WW, Kamen R, Weichselbaum R and Kufe D. . 1995 EMBO J. 14: 6148–6156.

  • Erhardt P, Tomaselli KJ and Cooper GM. . 1997 J. Biol. Chem. 272: 15049–15052.

  • Fanger GR, Gerwins P, Widmann C, Jarpe MB and Johnson GL. . 1997 Curr. Opin. Genet. Dev. 7: 67–74.

  • Goillot E, Raingeaud J, Ranger A, Tepper RI, Davis RJ, Harlow E and Sanchez I. . 1997 Proc. Natl. Acad. Sci. USA 94: 3302–3307.

  • Graves JD, Gotoh Y, Draves K, Ambrose D, Han DKM, Wright M, Chernoff J, Clark EA and Krebs EG. . 1998 EMBO J. 17: 2224–2234.

  • Hu MC-T, Qiu WR, Wang X, Meyer CF and Tan T-H. . 1996 Genes & Dev. 10: 2251–2264.

  • Ichijo H, Nishida E, Irie K, ten Dijke P, Saitoh M, Moriguchi T, Takagi M, Matsumoto K, Miyazono K and Gotoh Y. . 1997 Science 275: 90–94.

  • Ip YT and Davis RJ. . 1998 Curr. Opin. Cell Biol. 10: 205–219.

  • Kasibhatla S, Brunner T, Genestier L, Echeverri F, Mahboubi A and Green DR. . 1998 Mol. Cell 1: 543–551.

  • Kiefer F, Tibbles LA, Anafi M, Janssen A, Zanke BW, Lassam N, Pawson T, Woodgett JR and Iscove NN. . 1996 EMBO J. 15: 7013–7025.

  • Kothakota S, Azuma T, Reinhard C, Klippel A, Tang J, Chu K, McGarry TJ, Kirschner MW, Koths K, Kwiatkowski DJ and Williams LT. . 1997 Science 278: 294–298.

  • Kyriakis JM and Avruch J. . 1996 J. Biol. Chem. 271: 24313–24316.

  • Lazebnik YA, Kaufmann SH, Desnoyers S, Poirier GG and Earnshaw WC. . 1994 Nature 371: 346–347.

  • Lazebnik YA, Takahashi RD, Moir RD, Goldman RD, Poirier GG, Kaufmann SH and Earnshaw WC. . 1995 Proc. Natl. Acad. Sci. USA 92: 9042–9046.

  • Lenczowski JM, Lourdes D, Eder AM, King LB, Zacharchuk CM and Ashwell JD. . 1997 Mol. Cell. Biol. 17: 170–181.

  • Levkau B, Koyama H, Raines EW, Clurman BE, Herren B, Orth K and Roberts JM. . 1998 Mol. Cell 1: 553–563.

  • Ling P, Yao Z, Meyer CF, Wang XS, Oehrl W, Feller SM and Tan T-H. . 1999 Mol. Cell. Biol. 19: 1359–1368.

  • Meyer CF, Wang X, Chang C and Tan T-H. . 1996 J. Biol. Chem. 271: 8971–8976.

  • Oehrl W, Kardinal C, Ruf S, Adermann K, Groffen J, Feng G-S, Blenis J, Tan T-H and Feller SM. . 1998 Oncogene 17: 1893–1902.

  • Park DS, Stefanis L, Irene Yan CY, Farinelli SE and Greene LA. . 1996 J. Biol. Chem. 271: 21898–21905.

  • Pombo CM, Kehrl JH, Irma S, Woodgett JR, Force T and Kyriakis JM. . 1995 Nature 377: 750–754.

  • Rehemtulla A, Hamilton CA, Chinnaiyan AM and Dixit VM. . 1997 J. Biol. Chem. 272: 25783–25786.

  • Rudel T and Bokoch GM. . 1997 Science 276: 1571–1574.

  • Salvesen G and Dixit VM. . 1997 Cell 91: 443–446.

  • Seimiya H, Mashima T, Toha M and Tsuruo T. . 1997 J. Biol. Chem. 272: 4631–4636.

  • Shiah S-G, Chuang S-E, Chau Y-P, Shen S-C and Kuo M-L. . 1998 Cancer Res. 59: 391–398.

  • Song O, Lees-Miller SP, Kumar S, Zhang N, Chan DW, Smith GCM, Jackson SP, Alnemri ES, Litwack G, Khanna KK and Lavin MF. . 1996 EMBO J. 15: 3238–3246.

  • Steller H. . 1995 Science 267: 1445–1449.

  • Thompson CB. . 1995 Science 267: 1456–1462.

  • Tournier C, Whitmarsh AJ, Vavanagh J, Barret T and Davis RJ. . 1997 Proc. Natl. Acad. Sci. USA 94: 7337–7342.

  • Verheij M, Bose R, Lin XH, Yao B, Jarvis WD, Grant S, Birrer MJ, Szabo E, Zon LI, Kyriakis JM, Haimovitz-Friedman A, Fuks Z and Kolesnick RN. . 1996 Nature 380: 75–79.

  • Wang W, Zhou G, Hu MC-T, Yao Z and Tan T-H. . 1997 J. Biol. Chem. 272: 22771–22775.

  • Wen L-P, Fahrni JA, Troie S, Guan J-L, Orth K and Rosen GD. . 1997 J. Biol. Chem. 272: 26056–26061.

  • Widmann C, Gerwins P, Johson NL, Jarpe MB and Johnson GL. . 1998 Mol. Cell. Biol. 18: 2416–2429.

  • Xia Z, Dickens M, Raingeaud J, Davis RJ and Greenberg ME. . 1995 Science 270: 1326–1331.

  • Yao Z, Brown A, Wang XS, Zhou G, Diener K, Zukowski M and Tan T-H. . 1997a J. Biol. Chem. 272: 2118–2125.

  • Yao Z, Diener K, Wang XS, Zukowski M, Matsumoto G, Zhou G, Tibbles LA, Sasaki T, Nishina H, Tan T-H, Woodgett J and Penninger JM. . 1997b J. Biol. Chem. 272: 32378–32383.

  • Zanke BW, Boudreau K, Rubie E, Winnett E, Tibbles LA, Zon L, Kyriakis J, Liu F-F and Woodgett JR. . 1996 Curr. Biol. 6: 606–613.

  • Zhou G, Lee SC, Yao Z and Tan T-H. . 1999 J. Biol. Chem. 274: 13133–13138.

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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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