Peer-Reviewed Journal Details
Mandatory Fields
Fabian, Z,O'Brien, P,Pajecka, K,Fearnhead, HO
2009
October
Apoptosis
TPCK-induced apoptosis and labelling of the largest subunit of RNA polymerase II in Jurkat cells
Published
()
Optional Fields
TPCK Anti-tosyl antibody DFF40 DFF35/45 RNA polymerase II RPB1 CARBOXY-TERMINAL DOMAIN TOSYLPHENYLALANINE CHLOROMETHYL KETONE ELONGATION FACTOR-TU PRE-MESSENGER-RNA NF-KAPPA-B PROTEIN-SYNTHESIS NITROGEN-MUSTARD DNA FRAGMENTATION LEUKEMIC-CELLS ALPHA
14
1154
1164
N-alpha-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.
10.1007/s10495-009-0386-y
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