TPCK-induced apoptosis and labelling of the largest subunit of RNA polymerase II in Jurkat cells

Z. Fabian, P. O'Brien, K. Pajęcka, H. O. Fearnhead

Research output: Contribution to a Journal (Peer & Non Peer)Articlepeer-review

10 Citations (Scopus)

Abstract

N-α-Tosyl-l-phenylalanyl chloromethyl ketone (TPCK) is an affinity label for chymotrypsin-like proteases and has been extensively used as an experimental tool in apoptosis research to probe the role of proteases in cell death. While TPCK blocks some apoptotic changes and induces others, the cellular target or targets for TPCK have not been identified. Here we investigated for the first time the cellular targets for TPCK using a polyclonal anti-tosyl antibody. We have found that TPCK rapidly and irreversibly labels numerous intracellular proteins and have identified one as RPB1, the largest subunit of RNA polymerase II. We show that TPCK inhibits DNA binding by RNA polymerase and that TPCK inhibits transcription. Inhibition of transcription is known to induce apoptosis and while TPCK may trigger death through interaction with multiple targets, our data suggests that the pro-apoptotic effects of TPCK may be explained in part by the inhibition of RNA polymerase II activity.

Original languageEnglish
Pages (from-to)1154-1164
Number of pages11
JournalApoptosis
Volume14
Issue number10
DOIs
Publication statusPublished - Oct 2009
Externally publishedYes

Keywords

  • Anti-tosyl antibody
  • DFF35/45
  • DFF40
  • RNA polymerase II
  • RPB1
  • TPCK

Fingerprint

Dive into the research topics of 'TPCK-induced apoptosis and labelling of the largest subunit of RNA polymerase II in Jurkat cells'. Together they form a unique fingerprint.

Cite this