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J. Biol. Chem., Vol. 276, Issue 1, 827-834, January 5, 2001
From the Department of Molecular Biology, Princeton University,
Princeton, New Jersey 08544-1014
The human adenovirus type 2 E2 early (E2E)
transcriptional control region contains an efficient RNA polymerase III
promoter, in addition to the well characterized promoter for RNA
polymerase II. To determine whether this promoter includes intragenic
sequences, we examined the effects of precise substitutions introduced
between positions +2 and +62 on E2E transcription in an RNA polymerase III-specific, in vitro system. Two noncontiguous sequences
within this region were necessary for efficient or accurate
transcription by this enzyme. The sequence and properties of the
functional element proximal to the sites of initiation identified it as
an A box. Although a B box sequence could not be unambiguously located, substitutions between positions +42 and +62 that severely impaired transcription also inhibited binding of the human general initiation protein TFIIIC. Thus, this region of the RNA polymerase III E2E promoter contains a B box sequence. We also identified previously unrecognized intragenic sequences of the E2E RNA polymerase II promoter. In conjunction with our previous observations, these data
establish that RNA polymerase II and RNA polymerase III promoter sequences are superimposed from approximately positions -30 to +20 of
the complex E2E transcriptional control region. The alterations in
transcription induced by certain mutations suggest that components of
the RNA polymerase II and RNA polymerase III transcriptional machines
compete for access to overlapping binding sites in the E2E template.
Supported by a fellowship from the Medical Research Council of
Canada. Present address: University of Colorado Health Sciences Center,
Pediatric Hematology-Oncology Laboratories, Dept. of Pediatrics, 4200 East Ninth Ave., Denver, CO 80262.
§
To whom correspondence should be addressed: Tel.: 609-258-6113;
Fax: 609-258-2759; E-mail: sjflint@molbio.princeton.edu.
Copyright © 2001 by The American Society for Biochemistry and Molecular Biology, Inc. This article has been cited by other articles:
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