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J. Biol. Chem., Vol. 255, Issue 20, 9564-9570, 10, 1980
UM Hansen and WR McClure
The sigma subunit of Escherichia coli RNA polymerase was released from a
transcribing polymerase-poly[d(A-T)]-poly[d(A-T)]-RNA complex when the
nascent RNA reached a length of either eight or nine nucleotides. The
dissociated sigma was separated from other reaction components by gel
filtration, and was assayed using an activity assay previously described
(Hansen, U. M., and McClure, W. R. (1979) J. Biol. Chem. 254, 5713-5717).
The extent of RNA synthesis was limited by incorporation of 3'-dATP into
the RNA chains; a series of distributions of product lengths was achieved
by varying the concentration of 3'-dATP. A correlation of the number of
moles of dissociated sigma versus the number of moles of RNA products of
varying lengths allowed the determination of the point of release of the
sigma subunit. Models to explain the cause of sigma release are discussed.
Role of the sigma subunit of Escherichia coli RNA polymerase in initiation. II. Release of sigma from ternary complexes
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