HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS		QUICK SEARCH:   [advanced] Author: Keyword(s): Year:  Vol:  Page:

This Article Full Text Full Text (PDF) Alert me when this article is cited Alert me if a correction is posted Citation Map Services Email this article to a friend Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Jans, D. A. Articles by Trapani, J. A. Search for Related Content PubMed PubMed Citation Articles by Jans, D. A. Articles by Trapani, J. A. Social Bookmarking                      What's this?

Volume 271, Number 48, Issue of November 29, 1996 pp. 30781-30789
©1996 by The American Society for Biochemistry and Molecular Biology, Inc.

---

Nuclear Transport of Granzyme B (Fragmentin-2)
DEPENDENCE ON PERFORIN IN VIVO AND CYTOSOLIC FACTORS IN VITRO

(Received for publication, June 12, 1996, and in revised form, August 12, 1996)

David A. Jans , Patricia Jans , Lyndall J. Briggs , Vivien Sutton ^¶ and Joseph A. Trapani ^¶

From the  Nuclear Signalling Laboratory, Division for Biochemistry and Molecular Biology, John Curtin School of Medical Research, Canberra, Australian Capital Territory 2601 and ^¶ Cellular Cytotoxicity Laboratory, The Austin Research Institute, Heidelberg, Victoria 3084, Australia

Cytotoxic T and natural killer cells are able to kill their target cells through synergistic action of the pore-forming protein perforin and the serine protease granzyme B, resulting in very distinctive nuclear changes typical of apoptosis. Whereas perforin acts at the membrane, granzyme B appears to be both capable of entering the cytoplasm of target cells and accumulating in isolated nuclei. In this study we examine nuclear transport of fluoresceinated granzyme B both in vivo in intact cells in the presence of perforin and in vitro in semi-permeabilized cells using confocal laser scanning microscopy. Granzyme B alone was observed to enter the cytoplasm of intact cells but did not accumulate in nuclei. In the presence of sublytic concentrations of perforin, however, it accumulated strongly in intact cell nuclei to levels maximally about 1.5 times those in the cytoplasm after about 2.5 h. In vitro nuclear transport assays showed maximal levels of nuclear and nucleolar accumulation of granzyme B of about 2.5- and 3-fold those in the cytoplasm. In contrast to signal-dependent nuclear accumulation of SV40 large tumor antigen (T-Ag) fusion proteins in vitro, nuclear/nucleolar import of granzyme B was independent of ATP and not inhibitable by the non-hydrolyzable GTP analog GTPS (guanosine 5-O-(3-thiotriphosphate)). Similar to T-Ag fusion proteins, however, granzyme B nuclear and nucleolar accumulation was dependent on exogenously added cytosol. Specific inhibitors of granzyme B protease activity had no effect on nuclear/nucleolar accumulation, implying that proteolytic activity was not essential for nuclear targeting. The results imply that granzyme B (32 kDa) may be transported from the cytoplasm to the nucleus through passive diffusion and accumulate by binding to nuclear/nucleolar factors in a cytosolic factor-mediated process. Active and passive nuclear transport properties were normal in the presence of unlabeled granzyme B, implying that the nuclear envelope and pore complex are not granzyme B substrates.

CiteULike    Complore    Connotea    Del.icio.us    Digg    Reddit    Technorati    What's this?

This article has been cited by other articles:

				Y. Liu, L. H. Cheung, W. N. Hittelman, and M. G. Rosenblum Targeted delivery of human pro-apoptotic enzymes to tumor cells: In vitro studies describing a novel class of recombinant highly cytotoxic agents Mol. Cancer Ther., December 1, 2003; 2(12): 1341 - 1350. [Abstract] [Full Text] [PDF]

				R. N. Re Implications of intracrine hormone action for physiology and medicine Am J Physiol Heart Circ Physiol, March 1, 2003; 284(3): H751 - H757. [Full Text] [PDF]

				J. A. Trapani, V. R. Sutton, K. Y.T. Thia, Y. Q. Li, C. J. Froelich, D. A. Jans, M. S. Sandrin, and K. A. Browne A clathrin/dynamin- and mannose-6-phosphate receptor-independent pathway for granzyme B-induced cell death J. Cell Biol., January 21, 2003; 160(2): 223 - 233. [Abstract] [Full Text] [PDF]

				C. E. Hirst, M. S. Buzza, V. R. Sutton, J. A. Trapani, K. L. Loveland, and P. I. Bird Perforin-independent expression of granzyme B and proteinase inhibitor 9 in human testis and placenta suggests a role for granzyme B-mediated proteolysis in reproduction Mol. Hum. Reprod., December 1, 2001; 7(12): 1133 - 1142. [Abstract] [Full Text] [PDF]

				C. H. Bird, E. J. Blink, C. E. Hirst, M. S. Buzza, P. M. Steele, J. Sun, D. A. Jans, and P. I. Bird Nucleocytoplasmic Distribution of the Ovalbumin Serpin PI-9 Requires a Nonconventional Nuclear Import Pathway and the Export Factor Crm1 Mol. Cell. Biol., August 15, 2001; 21(16): 5396 - 5407. [Abstract] [Full Text] [PDF]

				M. J. Smyth, J. M. Kelly, V. R. Sutton, J. E. Davis, K. A. Browne, T. J. Sayers, and J. A. Trapani Unlocking the secrets of cytotoxic granule proteins J. Leukoc. Biol., July 1, 2001; 70(1): 18 - 29. [Abstract] [Full Text] [PDF]

				J. J. Yang, G. A. Preston, W. F. Pendergraft, M. Segelmark, P. Heeringa, S. L. Hogan, J. C. Jennette, and R. J. Falk Internalization of Proteinase 3 Is Concomitant with Endothelial Cell Apoptosis and Internalization of Myeloperoxidase with Generation of Intracellular Oxidants Am. J. Pathol., February 1, 2001; 158(2): 581 - 592. [Abstract] [Full Text] [PDF]

				V. R. Sutton, J. E. Davis, M. Cancilla, R. W. Johnstone, A. A. Ruefli, K. Sedelies, K. A. Browne, and J. A. Trapani Initiation of Apoptosis by Granzyme B Requires Direct Cleavage of Bid, but Not Direct Granzyme B-mediated Caspase Activation J. Exp. Med., November 13, 2000; 192(10): 1403 - 1414. [Abstract] [Full Text] [PDF]

				Y. Kawasaki, T. Saito, Y. Shirota-Someya, Y. Ikegami, H. Komano, M.-H. Lee, C. J. Froelich, N. Shinohara, and H. Takayama Cell Death-Associated Translocation of Plasma Membrane Components Induced by CTL J. Immunol., May 1, 2000; 164(9): 4641 - 4648. [Abstract] [Full Text] [PDF]

				C. Berthou, J.-F. Bourge, Y. Zhang, A. Soulie, D. Geromin, Y. Denizot, F. Sigaux, and M. Sasportes Interferon-gamma -induced membrane PAF-receptor expression confers tumor cell susceptibility to NK perforin-dependent lysis Blood, April 1, 2000; 95(7): 2329 - 2336. [Abstract] [Full Text] [PDF]

				K. A. Browne, E. Blink, V. R. Sutton, C. J. Froelich, D. A. Jans, and J. A. Trapani Cytosolic Delivery of Granzyme B by Bacterial Toxins: Evidence that Endosomal Disruption, in Addition to Transmembrane Pore Formation, Is an Important Function of Perforin Mol. Cell. Biol., December 1, 1999; 19(12): 8604 - 8615. [Abstract] [Full Text] [PDF]

				C. T. N. Pham and T. J. Ley Dipeptidyl peptidase I is required for the processing and activation of granzymes A and B in vivo PNAS, July 20, 1999; 96(15): 8627 - 8632. [Abstract] [Full Text] [PDF]

				K. L. Loveland, D. Herszfeld, B. Chu, E. Rames, E. Christy, L. J. Briggs, R. Shakri, D. M. de Kretser, and D. A. Jans Novel Low Molecular Weight Microtubule-associated Protein-2 Isoforms Contain a Functional Nuclear Localization Sequence J. Biol. Chem., July 2, 1999; 274(27): 19261 - 19268. [Abstract] [Full Text] [PDF]

				S. C. Piller, G. D. Ewart, D. A. Jans, P. W. Gage, and G. B. Cox The Amino-Terminal Region of Vpr from Human Immunodeficiency Virus Type 1 Forms Ion Channels and Kills Neurons J. Virol., May 1, 1999; 73(5): 4230 - 4238. [Abstract] [Full Text]

				D. A. Jans, V. R. Sutton, P. Jans, C. J. Froelich, and J. A. Trapani BCL-2 Blocks Perforin-induced Nuclear Translocation of Granzymes Concomitant with Protection against the Nuclear Events of Apoptosis J. Biol. Chem., February 12, 1999; 274(7): 3953 - 3961. [Abstract] [Full Text] [PDF]

				G. MacDonald, L. Shi, C. V. Velde, J. Lieberman, and A. H. Greenberg Mitochondria-dependent and -independent Regulation of Granzyme B-induced Apoptosis J. Exp. Med., January 4, 1999; 189(1): 131 - 144. [Abstract] [Full Text] [PDF]

				R. Hock, U. Scheer, and M. Bustin Chromosomal Proteins HMG-14 and HMG-17 Are Released from Mitotic Chromosomes and Imported into the Nucleus by Active Transport J. Cell Biol., December 14, 1998; 143(6): 1427 - 1436. [Abstract] [Full Text] [PDF]

				J. A. Trapani, D. A. Jans, P. J. Jans, M. J. Smyth, K. A. Browne, and V. R. Sutton Efficient Nuclear Targeting of Granzyme B and the Nuclear Consequences of Apoptosis Induced by Granzyme B and Perforin Are Caspase-dependent, but Cell Death Is Caspase-independent J. Biol. Chem., October 23, 1998; 273(43): 27934 - 27938. [Abstract] [Full Text] [PDF]

				J. L. Harris, E. P. Peterson, D. Hudig, N. A. Thornberry, and C. S. Craik Definition and Redesign of the Extended Substrate Specificity of Granzyme B J. Biol. Chem., October 16, 1998; 273(42): 27364 - 27373. [Abstract] [Full Text] [PDF]

				M. J. Pinkoski, M. Hobman, J. A. Heibein, K. Tomaselli, F. Li, P. Seth, C. J. Froelich, and R. C. Bleackley Entry and Trafficking of Granzyme B in Target Cells During Granzyme B-Perforin-Mediated Apoptosis Blood, August 1, 1998; 92(3): 1044 - 1054. [Abstract] [Full Text] [PDF]

				S. C. Piller, P. Jans, P. W. Gage, and D. A. Jans Extracellular HIV-1 virus protein R causes a large inward current and cell death in cultured hippocampal neurons: Implications for AIDS pathology PNAS, April 14, 1998; 95(8): 4595 - 4600. [Abstract] [Full Text] [PDF]

				D. Jans, L. Briggs, P Jans, C. Froelich, G Parasivam, S Kumar, V. Sutton, and J. Trapani Nuclear targeting of the serine protease granzyme A (fragmentin-1) J. Cell Sci., January 9, 1998; 111(17): 2645 - 2654. [Abstract] [PDF]

				W. Li, K. Nellaiappan, T. Strassmaier, L. Graham, K. M. Thomas, and H. M. Kagan Localization and activity of lysyl oxidase within nuclei of fibrogenic cells PNAS, November 25, 1997; 94(24): 12817 - 12822. [Abstract] [Full Text] [PDF]

				J. E. Niederhuber Cancer Vaccines: The Molecular Basis for T Cell Killing of Tumor Cells Oncologist, October 1, 1997; 2(5): 280 - 283. [Full Text] [PDF]

				A. Efthymiadis, H. Shao, S. Hubner, and D. A. Jans Kinetic Characterization of the Human Retinoblastoma Protein Bipartite Nuclear Localization Sequence (NLS) in Vivo and in Vitro. A COMPARISON WITH THE SV40 LARGE T-ANTIGEN NLS J. Biol. Chem., August 29, 1997; 272(35): 22134 - 22139. [Abstract] [Full Text] [PDF]

				K. A. Browne, R. W. Johnstone, D. A. Jans, and J. A. Trapani Filamin (280-kDa Actin-binding Protein) Is a Caspase Substrate and Is Also Cleaved Directly by the Cytotoxic T Lymphocyte Protease Granzyme B during Apoptosis J. Biol. Chem., December 8, 2000; 275(50): 39262 - 39266. [Abstract] [Full Text] [PDF]

				L. J. Schedlich, S. L. Le Page, S. M. Firth, L. J. Briggs, D. A. Jans, and R. C. Baxter Nuclear Import of Insulin-like Growth Factor-binding Protein-3 and -5 Is Mediated by the Importin beta Subunit J. Biol. Chem., July 28, 2000; 275(31): 23462 - 23470. [Abstract] [Full Text] [PDF]

				D. Zhang, M. S. Pasternack, P. J. Beresford, L. Wagner, A. H. Greenberg, and J. Lieberman Induction of Rapid Histone Degradation by the Cytotoxic T Lymphocyte Protease Granzyme A J. Biol. Chem., January 26, 2001; 276(5): 3683 - 3690. [Abstract] [Full Text] [PDF]