Requirement of the carboxy-terminal domain of RNA polymerase II for the transcriptional activation of chromosomal c-fos and hsp70A genes.
The carboxy-terminal domain of the large subunit of mouse and human RNA polymerase II contains 52 repeats of a heptapeptide which are the targets for a variety of kinases. We have used an alpha-amanitin resistant form of the large subunit of pol II to study the role of the carboxy-terminal domain in the expression of chromosomal genes. The large subunit of RNA polymerase II and deletion mutants thereof, which contain only 31 (LSdelta31) and 5 (LSdeltaS) repeats, were expressed in 293 cells. Subsequently, the endogenous large subunit of RNA polymerase II was inhibited by alpha-amanitin and the induction of chromosomal c-fos and hsp70A genes was determined. Cells expressing the large subunit of RNA polymerase II and LSdelta31 were able to transcribe the c-fos and hsp70A genes after treatment with the phorbolester TPA and after heat-shock, respectively. In contrast, cells expressing LSdelta5 failed to induce expression of both genes.
Meininghaus
M
M
GSF-Research Center for Environment and Health, Institute for Clinical Molecular Biology and Tumor Genetics, Munich, Germany.
Eick
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D000818:Animals; D002264:Carboxylic Acids; D002460:Cell Line; D016762:Genes, fos; D018840:HSP70 Heat-Shock Proteins / Q000235:genetics*; D006801:Humans; D051379:Mice; D012319:RNA Polymerase II / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D017384:Sequence Deletion; D015533:Transcriptional Activation
D002264:Carboxylic Acids; D018840:HSP70 Heat-Shock Proteins; D012319:RNA Polymerase II