Abstract
Publisher Summary Unlike most sequence-specific DNA recognition proteins, RNA polymerase (RNAP) participates in a series of molecular isomerizations subsequent to DNA binding. Determination of kinetic parameters by assaying products of abortive initiation takes advantage of the fact that RNAP does not translocate during the formation of the first several phosphodiester bonds in a nascent RNA chain. Differences between abortive and productive initiation kinetics can be used to examine changes in the frequency of natural abortive initiation as a result of mutation or the presence of an activator. Similarly, abortive initiation was used elegantly to dissect the role of purified protein factors in initiation of basal transcription in eukaryotes. In particular, it was possible to distinguish between factors necessary for formation of the first phosphodiester bond and those factors necessary for chain elongation.
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