Abstract
Lens epithelium-derived growth factor p75 (LEDGF/p75) is a nuclear autoantigen in atopic disorders implicated in cellular protection against stress-induced apoptosis. We observed that LEDGF/p75 was cleaved during apoptosis into fragments of 65 and 58 kD generated by caspases-3 and -7 cleaving at three sites: DEVPD30↓G, DAQD486↓G and WEID85↓N. Sequence analysis revealed that the DEVPD30↓G and WEID85↓N sites lie within the highly conserved HATH (homologous to amino terminus of hepatoma-derived growth factor) region, also known as PWWP domain. Alignment of proteins containing this domain failed to reveal conservation of the DEVPD30↓G and WEID85↓N sites, suggesting that the HATH/PWWP domain of LEDGF/p75 may be specifically targeted by caspases. Overexpression of LEDGF/p75 protected HepG2 cells from serum starvation-induced cell death, whereas expression of the 65 kD fragment failed to protect. The apoptotic cleavage of LEDGF/p75 may contribute to the pathogenesis of atopic disorders by abrogating its pro-survival function and enhancing its immunogenicity.
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Abbreviations
- DFS70:
-
dense fine speckled protein of 70 kD
- HATH:
-
homologous to amino terminal of hepatoma-derived growth factor
- HDGF:
-
hepatoma-derived growth factor
- HRP:
-
hepatoma-derived growth factor related protein
- LEDGF:
-
lens epithelium-derived growth factor
- PARP:
-
poly(ADP-ribose) polymerase
- PSI-BLAST:
-
Position Specific Iterated-Basic Local Alignment Search Tool
- PWWP:
-
proline-tryptophan-tryptophan-proline
- SDS–PAGE:
-
sodium dodecyl sulfate polyacrylamide gel electrophoresis
- STS:
-
staurosporine
- zVAD-fmk:
-
benzylozycarbonyl-Val-Ala-Asp-fluoromethylketone
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Acknowledgements
We are grateful to Drs Eng M Tan and Edward Chan (The Scripps Research Institute, La Jolla, CA, USA) and Dr Robert L Ochs (Precision Therapeutics, Pittsburgh, PA, USA) for valuable suggestions and discussions, for the kind gift of autoantibodies to LEDGF/p75, and for the pET28a-dfs70 plasmid. We also thank Drs Mark Johnson and Qinhong Ma (Loma Linda University) for valuable suggestions and discussions on sequence analysis, and Drs Marina Zemskova and Maria Filippova (Loma Linda University) for technical assistance. Support for microscopy and imaging facilities was provided by the Hedco Foundation. This work was supported by the National Institutes of Health Grant AI44088 to CA Casiano and by a Basic Science Research Grant from Loma Linda University School of Medicine.
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Wu, X., Daniels, T., Molinaro, C. et al. Caspase cleavage of the nuclear autoantigen LEDGF/p75 abrogates its pro-survival function: implications for autoimmunity in atopic disorders. Cell Death Differ 9, 915–925 (2002). https://doi.org/10.1038/sj.cdd.4401063
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DOI: https://doi.org/10.1038/sj.cdd.4401063
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