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Drosophila Factor 2, an RNA Polymerase II Transcript Release Factor, Has DNA-dependent ATPase Activity
Journal article   Open access   Peer reviewed

Drosophila Factor 2, an RNA Polymerase II Transcript Release Factor, Has DNA-dependent ATPase Activity

Zhi Xie and David Price
The Journal of biological chemistry, Vol.272(50), pp.31902-31907
12/12/1997
DOI: 10.1074/jbc.272.50.31902
PMID: 9395538
url
https://doi.org/10.1074/jbc.272.50.31902View
Published (Version of record) Open Access

Abstract

Drosophila factor 2 has been identified as a component of negative transcription elongation factor (N-TEF) that causes the release of RNA polymerase II transcripts in an ATP-dependent manner (Xie, Z. and Price D. H. (1996)J. Biol. Chem. 271, 11043–11046). We show here that the transcript release activity of factor 2 requires ATP or dATP and that adenosine 5′-O-(thiotriphosphate) (ATPγS), adenosine 5'-(β,γ-imino)triphosphate (AMP-PNP), or other NTPs do not support the activity. Factor 2 demonstrated a strong DNA-dependent ATPase activity that correlated with its transcript release activity. At 20 μg/ml DNA, the ATPase activity of factor 2 had an apparentKm(ATP) of 28 μm and an estimated Kcat of 140 min−1. Factor 2 caused the release of nascent transcripts associated with elongation complexes generated by RNA polymerase II on a dC-tailed template. Therefore, no other protein cofactors are required for the transcript release activity of factor 2. Using the dC-tailed template assay, it was found that renaturation of the template was required for factor 2 function.

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