Document Detail


TPCK-induced apoptosis and labelling of the largest subunit of RNA polymerase II in Jurkat cells.
MedLine Citation:
PMID:  19653099     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
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.
Authors:
Z Fabian; P O'Brien; K Pajecka; H O Fearnhead
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  Apoptosis : an international journal on programmed cell death     Volume:  14     ISSN:  1573-675X     ISO Abbreviation:  Apoptosis     Publication Date:  2009 Oct 
Date Detail:
Created Date:  2009-09-04     Completed Date:  2010-01-19     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  9712129     Medline TA:  Apoptosis     Country:  United States    
Other Details:
Languages:  eng     Pagination:  1154-64     Citation Subset:  IM    
Affiliation:
National Centre for Biomedical Engineering Science, School of Science, NUI Galway, Galway, Ireland.
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MeSH Terms
Descriptor/Qualifier:
Alkylation / drug effects
Antibodies / pharmacology
Apoptosis / drug effects*
Caspase 3 / metabolism
DNA / metabolism
DNA Fragmentation / drug effects
Deoxyribonucleases / metabolism
Enzyme Activation / drug effects
Humans
Jurkat Cells
Protein Binding / drug effects
Proteins / metabolism
RNA Polymerase II / metabolism*
Staining and Labeling*
Tosylphenylalanyl Chloromethyl Ketone / chemistry,  pharmacology*
Transcription, Genetic / drug effects
Chemical
Reg. No./Substance:
0/Antibodies; 0/DNA fragmentation factor, human; 0/Proteins; 402-71-1/Tosylphenylalanyl Chloromethyl Ketone; 9007-49-2/DNA; EC 2.7.7.-/RNA Polymerase II; EC 2.7.7.-/RNA polymerase II largest subunit; EC 3.1.-/DFFB protein, human; EC 3.1.-/Deoxyribonucleases; EC 3.4.22.-/Caspase 3

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine


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