Abstract
In this study, we evaluated the possible cross-talk between glucocorticoid (GC)-induced leucine zipper (Gilz) and caspase-8 in dexamethasone (Dex)-treated thymocytes. We determined that expression of Dex-induced Gilz protein was reduced when caspase-8 activity was inhibited, and this effect was not partially due to altered Gilz mRNA expression. Inhibition of the proteasome abrogated this reduction in Gilz expression, suggesting that Dex-induced caspase-8 activation protects Gilz from degradation. We hypothesized that the caspase-8-dependent protection of Gilz could be due to caspase-8-driven sumoylation. As a putative small ubiquitin-like modifier (SUMO)-binding site was identified in the Gilz sequence, we assessed whether SUMO-1 interacted with Gilz. We identified a 30-kDa protein that was compatible with the size of a GilzāSUMO-1 complex and was recognized by the anti-SUMO-1 and anti-Gilz antibodies. In addition, Gilz bound to SUMO ubiquitin-conjugating (E2)-conjugating enzyme Ube21 (Ubc9), the specific SUMO-1 E2-conjugating enzyme, in vitro and coimmunoprecipitated with Ubc9 in vivo. Furthermore, Gilz coimmunoprecipitated with SUMO-1 both in vitro and in vivo, and this interaction depended on caspase-8 activation. This requirement for caspase-8 was further evaluated in caspase-8-deficient thymocytes and lymphocytes in which Gilz expression was reduced. In summary, our results suggest that caspase-8 activation protects Gilz from proteasomal degradation and induces its binding to SUMO-1 in GC-treated thymocytes.
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Abbreviations
- GC:
-
glucocorticoid
- Gilz:
-
glucocorticoid-induced leucine zipper
- Dex:
-
dexamethasone
- TCR:
-
T-cell receptor
- GR:
-
glucocorticoid receptor
- GRE:
-
glucocorticoid-responsive element
- FoxO3:
-
Forkhead Box class O3
- p160 TIF2/GRIP1:
-
p160 cofactor transcriptional intermediary factor 2
- ENaC:
-
epithelial Na+ channel
- TAT:
-
transactivator of transcription
- E1:
-
ubiquitin-activating enzyme
- E2:
-
ubiquitin-conjugating enzyme
- E3:
-
ubiquitin ligase
- UBL:
-
ubiquitin-like protein
- SUMO:
-
small ubiquitin-like modifier
- Ubc9:
-
SUMO E2-conjugating enzyme Ube21
- Gilz-GST:
-
Gilz bound to GST
- PHA:
-
phytohemagglutinin
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Acknowledgements
This work was supported in part by Associazione Italiana Ricerca Cancro (AIRC) and by the Italian Ministry of University (PRIN 2008RFPZN7_002).
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Delfino, D., Spinicelli, S., Pozzesi, N. et al. Glucocorticoid-induced activation of caspase-8 protects the glucocorticoid-induced protein Gilz from proteasomal degradation and induces its binding to SUMO-1 in murine thymocytes. Cell Death Differ 18, 183ā190 (2011). https://doi.org/10.1038/cdd.2010.86
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DOI: https://doi.org/10.1038/cdd.2010.86
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