Skip to main content
construction release_alert
Scholars@Duke will be undergoing maintenance April 11-15. Some features may be unavailable during this time.
cancel

Evidence that non-caspase proteases are required for chromatin degradation during apoptosis.

Publication ,  Journal Article
Hughes, FM; Evans-Storms, RB; Cidlowski, JA
Published in: Cell Death Differ
December 1998

Chromatin degradation into oligonucleosomal and approximately 30-50 Kb fragments is a hallmark of apoptosis. Crude nuclear extract from apoptotic rat thymocytes is able to recapitulate both types of DNA fragmentation in an assay using HeLa cell nuclei as an exogenous substrate. Using size exclusion chromatography we have identified a novel activity (approximately 260 Kd) that produces only approximately 30-50 Kb DNA fragments, and a 25 Kd activity that generates both approximately 30-50 Kb and oligonucleosomal fragments. Both activities produced DNA fragments with 3'-OH termini, are dependent on Ca2+ and Mg2+ and are inhibited by N-ethyl-maleimide, sodium tetrathionate, aurintricarboxylic acid and sodium chloride, similar to other nucleases implicated in apoptosis. These activities were inhibited by the serine protease inhibitors N-tosyl-L-phenylalanine chloromethyl ketone and N alpha-p-tosyl-L-lysine chloromethyl ketone, but not by the serine protease inhibitor diisopropyl fluorophosphate, or by calpain inhibitors I or II, or the capsase inhibitors Ac-Asp-Glu-Val-Asp-aldehyde, Ac-Tyr-Val-Ala-Asp-aldehyde, or Z-Val-Ala-Asp-fluoromethyl ketone. Both activities were insensitive to protease inhibitors when extracts were incubated with naked linear DNA, indicating the presence of both nuclease and protease activities in the preparation. Together, these observations suggest the involvement of non-caspase proteases in apoptosis which perhaps function by altering chromatin substructure and exposing it to nucleolytic attack.

Duke Scholars

Published In

Cell Death Differ

DOI

ISSN

1350-9047

Publication Date

December 1998

Volume

5

Issue

12

Start / End Page

1017 / 1027

Location

England

Related Subject Headings

  • Tosylphenylalanyl Chloromethyl Ketone
  • Tosyllysine Chloromethyl Ketone
  • Thymus Gland
  • Serine Proteinase Inhibitors
  • Serine Endopeptidases
  • Rats, Sprague-Dawley
  • Rats
  • Protease Inhibitors
  • Peptide Fragments
  • Oligopeptides
 

Citation

APA
Chicago
ICMJE
MLA
NLM
Hughes, F. M., Evans-Storms, R. B., & Cidlowski, J. A. (1998). Evidence that non-caspase proteases are required for chromatin degradation during apoptosis. Cell Death Differ, 5(12), 1017–1027. https://doi.org/10.1038/sj.cdd.4400418
Hughes, F. M., R. B. Evans-Storms, and J. A. Cidlowski. “Evidence that non-caspase proteases are required for chromatin degradation during apoptosis.Cell Death Differ 5, no. 12 (December 1998): 1017–27. https://doi.org/10.1038/sj.cdd.4400418.
Hughes FM, Evans-Storms RB, Cidlowski JA. Evidence that non-caspase proteases are required for chromatin degradation during apoptosis. Cell Death Differ. 1998 Dec;5(12):1017–27.
Hughes, F. M., et al. “Evidence that non-caspase proteases are required for chromatin degradation during apoptosis.Cell Death Differ, vol. 5, no. 12, Dec. 1998, pp. 1017–27. Pubmed, doi:10.1038/sj.cdd.4400418.
Hughes FM, Evans-Storms RB, Cidlowski JA. Evidence that non-caspase proteases are required for chromatin degradation during apoptosis. Cell Death Differ. 1998 Dec;5(12):1017–1027.

Published In

Cell Death Differ

DOI

ISSN

1350-9047

Publication Date

December 1998

Volume

5

Issue

12

Start / End Page

1017 / 1027

Location

England

Related Subject Headings

  • Tosylphenylalanyl Chloromethyl Ketone
  • Tosyllysine Chloromethyl Ketone
  • Thymus Gland
  • Serine Proteinase Inhibitors
  • Serine Endopeptidases
  • Rats, Sprague-Dawley
  • Rats
  • Protease Inhibitors
  • Peptide Fragments
  • Oligopeptides