Open AccessTranscription of Yeast DNA in vitro
Author(s) -
PONTA Helmut,
PONTA Ulrike,
KRAFT Volker,
WINTERSBERGER Erhard
Publication year - 1974
Publication title -
european journal of biochemistry
Language(s) - English
Resource type - Journals
eISSN - 1432-1033
pISSN - 0014-2956
DOI - 10.1111/j.1432-1033.1974.tb03640.x
Subject(s) - biochemistry , transcription (linguistics) , microbiology and biotechnology , yeast , dna , rna , polymerase , gtp' , termination factor , nucleotide , biology , chemistry , rna dependent rna polymerase , enzyme , gene , philosophy , linguistics
Yeast DNA as prepared by usual procedures is shown to contain a very effective inhibitor of the initiation of the transcription in vitro. The presence of this inhibitor is responsible for the low template activity of yeast DNA observed by several workers. After removal of the inhibitor by CsCl gradient centrifugation, yeast DNA is readily transcribed by the purified RNA polymerases A and B. Some of the characteristics of the transcription reaction such as the effects of divalent cation (Mn 2+ and Mg 2+ ) and salt are described. The purine nucleotide used for initiation of RNA chains was determined. While RNA polymerase A initiates exclusively with ATP RNA polymerase B uses both GTP and ATP for initiation but strongly prefers GTP. Preliminary studies on the transcription inhibitor removed from yeast DNA preparations by CsCl gradient centrifugation suggest that this substance is polyphosphate.