Research Article: The Death Effector Domains of Caspase-8 Induce Terminal Differentiation

Date Published: November 18, 2009

Publisher: Public Library of Science

Author(s): Ainhoa Mielgo, Vicente A. Torres, Michael C. Schmid, Ryon Graf, Samantha G. Zeitlin, Pedro Lee, David J. Shields, Simone Barbero, Colin Jamora, Dwayne G. Stupack, Mikhail V. Blagosklonny. http://doi.org/10.1371/journal.pone.0007879

Abstract: The differentiation and senescence programs of metazoans play key roles in regulating normal development and preventing aberrant cell proliferation, such as cancer. These programs are intimately associated with both the mitotic and apoptotic pathways. Caspase-8 is an apical apoptotic initiator that has recently been appreciated to coordinate non-apoptotic roles in the cell. Most of these functions are attributed to the catalytic domain, however, the amino-terminal death effector domains (DED)s, which belong to the death domain superfamily of proteins, can also play key roles during development. Here we describe a novel role for caspase-8 DEDs in regulating cell differentiation and senescence. Caspase-8 DEDs accumulate during terminal differentiation and senescence of epithelial, endothelial and myeloid cells; genetic deletion or shRNA suppression of caspase-8 disrupts cell differentiation, while re-expression of DEDs rescues this phenotype. Among caspase-8 deficient neuroblastoma cells, DED expression attenuated tumor growth in vivo and proliferation in vitro via disruption of mitosis and cytokinesis, resulting in upregulation of p53 and induction of differentiation markers. These events occur independent of caspase-8 catalytic activity, but require a critical lysine (K156) in a microtubule-binding motif in the second DED domain. The results demonstrate a new function for the DEDs of caspase-8, and describe an unexpected mechanism that contributes to cell differentiation and senescence.

Partial Text: Caspase-8 is an initiator protease recruited to the death inducing signaling complex during apoptosis initiated by death receptors. Homotypic interactions, mediated by the amino-terminal death effector domains (DEDs) of caspase-8, are required for recruitment and subsequent maturation and dimerization of caspase-8 initiating the extrinsic apoptosis cascade. In addition to this role in death receptor-mediated apoptosis, cumulative evidence suggests that caspase-8 performs other non-apoptotic functions in development [1], including proliferation [2], [3], cell migration [4]–[7] and differentiation [3], [8]. We and others previously reported that caspase-8 has the capacity to localize to a number of different cellular locations, including the cytosolic compartment [9], actin-rich ruffles [10], [11], endosomes [12], including those at the front of migrating cells[13], focal adhesions [4] and stable microtubule structures, such as centrosomes [14]. Interestingly, the different domains of caspase-8 appear to favor localization to different cellular compartments. It is possible that these different preferred locations may ultimately influence caspase-8 function(s).

In this study, we characterize a new function for the death effector domains of caspase-8 in cell cycle regulation. First, we find that caspase-8 DEDs accumulate in cells undergoing terminal differentiation. Silencing of caspase-8 disrupts or delays differentiation, while reintroduction of DEDs restores differentiation potential. We further report that DED expression is sufficient to impair tumor growth and cell proliferation, promoting mitotic defects that foster cell death or cell cycle arrest and terminal differentiation. Finally, we demonstrate that these events require a critical lysine (K156) in a microtubule binding motif in the second DED (DED-b). These observations together suggest that caspase-8 DEDs function as a tumor suppressor, acting as an anti-proliferation and differentiation-inducing element. It is tempting to speculate that this type of mechanism may act in some spontaneously regressing tumors, such as stage IV-S neuroblastoma, however, such a mechanism would be co-dependent upon other classic tumor suppression pathways, including p53 and p21. Interestingly, the cell lines which can maintain higher levels of DEDs, such as COS-7, also have documented defects in p53 signaling.

Source:

http://doi.org/10.1371/journal.pone.0007879

 

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