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Dissociation and reconstitution of active DNA‐dependent RNA‐polymerase from E. coli
Author(s) -
Sethi V.Sagar,
Zillig W.,
Bauer H.
Publication year - 1970
Publication title -
febs letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.593
H-Index - 257
eISSN - 1873-3468
pISSN - 0014-5793
DOI - 10.1016/0014-5793(70)80274-5
Subject(s) - citation , planck , physics , rna polymerase , transcription (linguistics) , dna , dna polymerase , microbiology and biotechnology , polymerase , philosophy , chemistry , rna , stereochemistry , biology , quantum mechanics , biochemistry , library science , linguistics , gene , computer science
DNA-dependent RNA-polymerase from E.coli consists of 4-5 subunits, namely /3’ (MW 160,000), /I (145,000), u (85,000), (Y (40,000) and may be w (12,000) [ 1,2]. These have been separated by column chromatography on Sephadex G-200 in 1% SDS and on DEAE-cellulose in 8 M urea [2] as well as by preparative electrophoresis on cellulose acetate blocks in 6 M urea [ 11. Attempts to reconstitute active enzyme from these subunits have been unsuccessful so far. Ishihama and Hurwitz [3] reported a partial recovery of enzymatic activity after incubation with DNA of fragments which had been obtained by the dissociation of enzyme at low urea concentrations. Recently we reported the dissociation of enzyme into its subunits in 4 M lithium chloride and showed that the isolated sigma subunit was fully active in stimulating the activity of the core enzyme [4]. In this paper we report the reconstitution of the active enzyme from isolated subunits as well as the specific binding of the /3’/3-fraction to 3 H-labelled T4-DNA.