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Fractionation of transcription factors for RNA polymerase II from Drosophila Kc cell nuclear extracts
Journal article   Open access   Peer reviewed

Fractionation of transcription factors for RNA polymerase II from Drosophila Kc cell nuclear extracts

David H Price, Ann E Sluder and Arno L Greenleaf
The Journal of biological chemistry, Vol.262(7), pp.3244-3255
1987
DOI: 10.1016/S0021-9258(18)61497-8
PMID: 3818640
url
https://doi.org/10.1016/S0021-9258(18)61497-8View
Published (Version of record) Open Access

Abstract

Drosophila Kc cells were utilized to prepare nuclear extracts in which promoter-containing DNA templates were efficiently transcribed by RNA polymerase II. A combination of fractionation schemes was used to identify and partially purify seven activities (factors) which affected the transcription of four different genes in vitro. Reconstructing specific transcription required exogenous RNA polymerase II in addition to these factors. Moreover, the high efficiency of transcription characteristic of the crude extract was preserved in reconstruction reactions. The methods used are presented in detail. Functions were assigned to several of the factors. One essential factor appeared to affect initiation and displayed chromatographic properties unlike any other Drosophila transcription factor previously described. Two factors specifically affected RNA chain elongation. Another activity was a DNase inhibitor required to preserve template integrity in the fractionated system. The remaining three factors were not absolutely essential but affected the specific in vitro transcription either qualitatively or quantitatively. A comparison of these transcription factors with other Drosophila and mammalian transcription factors is made.
 Fundamental and applied biological sciences. Psychology Biological and medical sciences Analytical, structural and metabolic biochemistry Enzymes and enzyme inhibitors Transferases

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