Abstract
Special AT-rich sequence-binding protein 1 (SATB1), predominantly expressed in thymocytes, was identified as a component of the nuclear matrix protein fraction. Programmed cell death of Jurkat T-cells was induced by various stimuli in Fas-dependent and -independent fashion. During apoptosis, but not during necrosis, SATB1 was cleaved, as rapidly as was lamin B, in a caspase-dependent way yielding a stable 70 kDa fragment. The same result was obtained for apoptotic HL60-cells. We constructed various deletion constructs of SATB1, expressing protein chimeras tagged with green fluorescent protein (GFP). Transient transfection of these into Jurkat or HeLa cells followed by initiation of apoptosis allowed us to map the potential caspase-6 cleavage site VEMD to the N-terminal third of SATB1, leaving an intact DNA-binding domain in the C-terminal part of the protein. Our results suggest that apoptosis-specific breakdown of SATB1, a transcriptional activator of the CD8a gene, might be of physiological relevance during thymic clonal deletion and apoptosis of peripheral T-lymphoid cells. Cell Death and Differentiation (2000) 7, 425–438
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Abbreviations
- SATB1:
-
special AT-rich binding protein 1
- M/SAR:
-
matrix/scaffold attachment region
- NMPs:
-
nuclear matrix proteins
- GFP:
-
green fluorescent protein
- PCD:
-
programmed cell death
- -fmk:
-
-fluoromethylketone
- -cmk:
-
-chloromethylketone
- -CHO:
-
-aldehyde
- TLCK:
-
N-α-p-tosyl-L-lysine chloromethylketone
- TPCK:
-
N-tosyl-L-phenylalanine-chloromethylketone
- fl:
-
full length
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Acknowledgements
The authors wish to thank Terumi Kohwi-Shigematsu (University of California, Berkeley, CA, USA) for the generous gift of anti-SATB1 serum, and are indebted to Roland Foisner (Vienna Biocenter, Austria) for kindly providing anti-nucleolin (3G4B2) antibody. We highly appreciate fruitful discussions with Wilfried Bursch, Rolf Schulte-Hermann and Georg Sauermann and wish to thank the latter two for critical reading of the manuscript. Furthermore, we wish to thank Dieter Gelbmann and Editha Bayer for technical assistance and Ulrike Fröhwein for help with cell culture. This work was supported by a grant from the Herzfelder Stiftung, Vienna, Austria.
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Gotzmann, J., Meissner, M. & Gerner, C. The fate of the nuclear matrix-associated-region-binding protein SATB1 during apoptosis. Cell Death Differ 7, 425–438 (2000). https://doi.org/10.1038/sj.cdd.4400668
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