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DNA dependent RNA polymerase from rat liver mitochondria
Author(s) -
Gallerani R.,
Saccone C.,
Cantatore P.,
Gadaleta M.N.
Publication year - 1972
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(72)80213-8
Subject(s) - library science , chemistry , biological sciences , citation , computer science , biology , computational biology
The purification of mitochondrial DNA-dependent RNA polymerase from rat liver and other organisms appears to be particularly difficult since the enzyme is readily inactivated by a wide range of extraction procedures [ 1,2] . We recently succeeded in solubilizing the enzyme from rat liver mitochondria using the detergent deoxycholate [3]. Stabilization of the enzyme activity after extraction was achieved with large concentrations of dithiothreitol and glycerol [3]. Mitochondrial rat liver RNA polymerase solubilized either by sonication or by deoxycholate ap pears to belong to the bacterial class of RNA polymerases since is inhibited by rifampicin and insensitive to cw-amanitin [3,4] . A mitochondrial RNA polymerase has recently been purified from sonicates of Neurospora mitochondria by repeated glycerol gradient centrifugation [5]. Solubilization and partial purification of mitochondrial enzyme from wild type yeast has been reported by Tsai et al. [6] . In this paper we report the first attempt to purify a mitochondrial RNA polymerase from animal cells. Partial purification of the enzyme was achieved by a procedure including lysis at high ionic strength, DNase treatment, ammonium sulphate fractionation and DEAE-Sephadex A-25 column chromatography.