Initiation and Release of RNA by DNA-Dependent RNA Polymerase

During in vitro transcription of T4 DNA by E. coli RNA polymerase, chain initiation stops coincidentally with synthesis at low ionic strength (0.11) with an average of one RNA chain initiated per 24S polymerase molecule. At high ionic strength (0.37), initiation as well as synthesis continues for several hours, with an average of four chains initiated per enzyme molecule in two hours. The RNA product is released from the T4 DNA template at both low and high ionic strength. At high ionic strength, however, RNA polymerase can repeatedly initiate, synthesize, and release RNA from the synthesis complex in vitro. Under both conditions, the synthesized RNA sediments at 25-44 S, has a number average chain length of 5000 to 7500 nucleotides, and a weight average chain length of 11,500 nucleotides.