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(Received for publication, August 22, 1995; and in revised form, November 15, 1995 ) The 40.6-kDa
Volume 271,
Number 5,
Issue of February 2, 1996 pp. 2482-2490
©1996 by The American Society for Biochemistry and Molecular Biology, Inc.
*,
a UV-inducible Smaller Form of the
Subunit Sliding Clamp of DNA
Polymerase III of Escherichia coli
I. GENE EXPRESSION AND REGULATION
subunit of DNA polymerase III of Escherichia coli is a sliding DNA clamp responsible for
tethering the polymerase to DNA and endowing it with high processivity
(Stukenberg, P. T., Studwell-Vaughan, P. S., and O'Donnell,
M.(1991) J. Biol. Chem. 266, 11328-11334). UV
irradiation of E. coli induces a smaller 26-kDa form of the
subunit, termed
*, that, when overproduced from a plasmid,
increases UV resistance of E. coli (Skaliter, R., Paz-Elizur,
T., and Livneh, Z.(1996) J. Biol. Chem. 271, 2478-2481).
Here we show that this protein is synthesized from a UV-inducible
internal gene, termed dnaN*, that is located in-frame inside
the coding region of dnaN, encoding the
subunit. The
initiation codon and the Shine-Dalgarno sequence of dnaN* were
identified by site-directed mutagenesis. The dnaN* transcript
was shown to be induced upon treatment with nalidixic acid, and
transcriptional dnaN*-cat gene fusions were UV inducible,
suggesting induction of dnaN* at the transcriptional level.
Analysis of translational dnaN*-lacZ gene fusions revealed
that UV induction was abolished in strains carrying the recA56, lexA3, or
rpoH mutations,
indicating involvement of both SOS and heat shock stress responses in
the induction process. Expression of dnaN* represents a
strategy of producing several proteins with related functional domains
from a single gene.
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