Open AccessDrosophila TFIIE: Purification, cloning, and functional reconstitution
Author(s) -
Xia Wang,
Steven G. Hansen,
Ryan Ratts,
Shi-Sheng Zhou,
Amy J. Snook,
William A. Zehring
Publication year - 1997
Publication title -
proceedings of the national academy of sciences of the united states of america
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 5.011
H-Index - 771
eISSN - 1091-6490
pISSN - 0027-8424
DOI - 10.1073/pnas.94.2.433
Subject(s) - biology , complementary dna , transcription factor ii a , microbiology and biotechnology , transcription preinitiation complex , genetics , general transcription factor , polytene chromosome , molecular cloning , drosophila melanogaster , gene , gene expression , promoter
We present a physical and molecular genetic characterization ofDrosophila melanogaster TFIIE (dTFIIE), a component of the basal RNA polymerase II transcription apparatus. We have purified dTFIIE to near homogeneity from nuclear extracts ofDrosophila embryos and found that it is composed of two subunits with apparent molecular weights of 55 and 38 kDa. Peptide sequence information derived from the two subunits was used to isolate the corresponding cDNA clones, revealing that dTFIIE and human TFIIE share extensive amino acid similarity. Functional conservation was demonstrated by the ability of bacterially expressed dTFIIE to substitute for human TFIIE in anin vitro transcription assay reconstituted from purified components. Cytological mapping analysis shows that both subunits are encoded by single copy genes located on chromosome III.