# GENETIC AND BIOCHEMICAL ANALYSIS OF PROMOTOR FUNCTION

> **NIH NIH R01** · UNIVERSITY OF IOWA · 1990 · $145,248

## Abstract

Genetic (mutational) and biochemical methods will be used to
investigate mechanisms of transcription initiation and its
regulation in well-defined prokaryotic systems.  Two promoters
(transcription initiation signals) of bacteriophage lambdal will
be studied in order to address the following questions: a) How do
specific DNA sequences provide information for discrete steps in
transcription initiation? b) What is the nature of the interaction
between RNA polymerase and specific activator proteins that allows
the activators to stimulate transcription initiation from weak
promoters?  c) How does the DNA sequence with which the polymerase
interacts influence the interaction between the enzyme and a
specific activator bound to an adjacent or overlapping recognition
site?

The specific aims of the proposed research are: (1) Use of
oligonucleotide-directed mutagenesis to isolate specific mutations
in the lambdal PRM promoter and evaluation of the effects of the
mutations on promoter function in vitro. (2) Isolation of mutations
in the lambdal cI protein--the specific activator of PRM--that
would allow the protein to activate an altered promoter. (3)
Isolation of mutations in the lambdal cII protein--the specific
activator of PRE--that would affect the ability of the protein to
activate PRE without affecting its ability to bind to DNA. (4)
Kinetic analysis of the ability of lambdal cII protein to bind to
wild-type PRE and PRE mutant DNA's.  (5) Isolation of RNA
polymerase mutants with altered ability to be activated by cII or
cI protein.  (6) Kinetic characterization of interactions of mutant
proteins with their DNA recognition sites and with each other.

These studies should contribute to an understanding of regulatory
processes in higher organisms as well as in prokaryotes; they
should help elucidate the nature of DNA sequence elements required
for interaction with complex recognition proteins; and they should
provide insights into protein-protein interactions that regulate
transcription.  The proposed research will be especially relevant
to the ways that proteins interact at multiple regulatory sites in
higher organisms to control gene activity in response to external
stimuli.

## Key facts

- **NIH application ID:** 3127254
- **Project number:** 5R01AI017508-10
- **Recipient organization:** UNIVERSITY OF IOWA
- **SAM.gov UEI:** Z1H9VJS8NG16
- **Principal Investigator:** GARY N GUSSIN
- **Activity code:** R01 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 1990
- **Award amount:** $145,248
- **Award type:** 5
- **Project period:** 1980-12-01 → 1992-11-30

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/3127254

## Citation

> US National Institutes of Health, RePORTER application 3127254, GENETIC AND BIOCHEMICAL ANALYSIS OF PROMOTOR FUNCTION (5R01AI017508-10). Retrieved via MORVS 2026-10-25 from https://api.morvs.ai/grant/nih/3127254. Licensed CC0.

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