# CONTROL OF ADENOVIRUS TRANSCRIPTION

> **NIH NIH R01** · WISTAR INSTITUTE · 1988 · $169,000

## Abstract

The activation of oncogenic cellular genes involved in transformation and
of many other genes seems to be, at least in some cases, at the level of
transcription.  This proposal centers on the analysis of initiation of
transcription by RNA polymerases II and III in vitro systems which
faithfully reproduce the site and requirements for in vivo initiation of
transcription.  We will use a combination of straight biochemical
fractionation approaches with a genetic approach which takes advantage of
template and transcriptional machinery mutants as well as nucleotide
analogs.  The RNA polymerase II work will focus on the proteins involved in
initiation by analyzing in detail the biochemical (enzyme with cofactors)
as well as genetic (i.e., in the template sequence) features required for
the formation of the transcription initiation complex.  We will use
transcriptional extracts from HeLa cells and templates consisting of
oncogenic adenovirus DNA inserted in bacterial plasmids or phages.  Novel
features of the initiation process, like for example, unwinding of the DNA
double helix or the mechanism of action of the inhibitor of transcription
initiation, DRB, will be some of the partial processes that will be
analyzed in detail in transcriptional initiation complexes.  The unwinding
assay will also be used for the VAI RNA genes transcribed by RNA polymerase
III.  The formation of a transcriptional complex, which distinguishes
between the initiation and elongation phases of transcription, revealed a
large molar excess of initiation events.  This excess could be related to a
mechanism of gene activation by an increase in the amounts of elongation
factors or might be an artifact of the in vitro system.  Point mutants in
the DNA template will be generated by site-specific mutagenesis on M-13
cloning vectors which allow rapid sequence analysis of the mutants and in
vitro transcription with the replicative form of the recombinant DNA
phage.  Site-specific point mutants allow to better establish the role of
each element of the promoter sequence, since the environment around the
mutation remains constant.  The interactions between the mutant or
wild-type promoter sequences and the elements of the transcriptional
machinery, cellular RNA polymerase II Alpha-amanitin resistant mutants and
transcriptionally resistant DRB resistant mutants will be analyzed.
Mutations in the RNA polymerase or factors should produce some abnormal
interactions.  We will continue our efforts in establishing the mechanism
of action of the transcription initiation inhibitor DRB and the biochemical
basis for these cell mutants.

## Key facts

- **NIH application ID:** 3125417
- **Project number:** 5R01AI013231-11
- **Recipient organization:** WISTAR INSTITUTE
- **SAM.gov UEI:** DW1XZMGNFBL4
- **Principal Investigator:** ROBERTO WEINMANN
- **Activity code:** R01 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 1988
- **Award amount:** $169,000
- **Award type:** 5
- **Project period:** 1976-06-30 → 1989-06-30

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/3125417

## Citation

> US National Institutes of Health, RePORTER application 3125417, CONTROL OF ADENOVIRUS TRANSCRIPTION (5R01AI013231-11). Retrieved via MORVS 2026-10-25 from https://api.morvs.ai/grant/nih/3125417. Licensed CC0.

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