Abstract
1. 1. A simple procedure to obtain DNA-free RNA polymerase (nucleosidetriphosphate: RNA nucleotidyltransferase, EC 2.7.7.6) from HeLa cells was developed from the preparation of extracts in a hypotonic medium in a French pressure cell. 2. 2. The extracts contained two forms of RNA polymerase: ‘freed’ from, or ‘complexed’ to, DNA. Free enzyme was observed to consist of 10-S and 20-S RNA polymerase. The complex enzyme obtained here was supposed to correspond to a ‘specific complex’, in which all the bound enzymes are activated in preparation for the reaction. 3. 3. The partially purified preparations of HeLa RNA polymerase thus obtained exhibited similar reaction profiles to those of Escherichia coli RNA polymerase, except for the following two characteristics; one was the weak affinity for substrates and the other was the slow rate of RNA chain growth. 4. 4. Alterations in the intracellular distribution of the enzyme were surveyed in relation to growth states of the cultured cells. The biological significance was considered to reflect some regulations of gene expressions.
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