# STREPTOCOCCUS PLASMIDS--MOLECULAR GENETIC ANALYSIS

> **NIH NIH R01** · DUKE UNIVERSITY · 1993 · $234,717

## Abstract

The only approach to the control of bacterial infections is the use of
antibacterial drugs.  However, antibiotic resistance can compromise
treatment.  One of the most frequently used antibiotics is tetracycline.
The Tet M resistance determinant, which we discovered during the course
of this project, has proven to have an extremely broad host range among
bacterial pathogens.  We have shown that Tet(M) protein protects the
protein biosynthetic machinery of the cell from the inhibitory action of
antibiotic at the level of the ribosome.  The goal of this project is to
understand the molecular basis for this protection mechanism with the
hope that such information may be useful in design of new tetracycline
derivatives which may be useful in treating infections with a possible
secondary outcome being a better understand the mechanism of tetracycline
action.  To this end we plan to study the certain steps during protein
synthesis for their susceptibility to tetracycline and the consequence of
the presence of Tet(M) protein.  In particular preliminary experiments
suggest that Tet(M) protein may function as a homolog of the normal
elongation factor G (which functions during ribosome translocation). some
extent the approaches outlined below will be guided by this observation.

Two general approaches will be employed: (1) A biochemical approach will
be used to evaluate the physical interaction of Tet(M) with protein
biosynthetic components.  For example, we have shown that the protein has
high affinity for the ribosome, although it may also interact with
elongation factors.  The nature of such interactions will be examined by
use of protein-protein and protein-nucleic acid crosslinking reagents to
deduce interaction of Tet(M) with these components, and by application of
alkylation protection methods to assess rRNA interactions with
tetracycline in the presence and absence Tet(M). (2) These biochemical
experiments will be complemented by genetic analysis of Escherichia coli
chromosomal mutations that were isolated during the previous grant period
which confer tetracycline sensitivity on the cell even in the presence of
functional Tet(M).  Our current hypothesis is that such mutations alter
the gene(s) that encode tetracycline binding activities and/or components
which interact directly with Tet(M).  Preliminary mapping of these
mutations place them within the major cluster of ribosomal protein genes
(73 min).  We hope to clone and identify the corresponding genes and
characterize the nature of the mutations conferring this phenotype.  In a
related approach, we plan attempted isolation of mutations in the rRNA
coding sequences to see if we can identify alterations in these molecules
that result in tetracycline resistance.

## Key facts

- **NIH application ID:** 3126290
- **Project number:** 2R01AI015619-14
- **Recipient organization:** DUKE UNIVERSITY
- **SAM.gov UEI:** TP7EK8DZV6N5
- **Principal Investigator:** VICKERS BURDETT
- **Activity code:** R01 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 1993
- **Award amount:** $234,717
- **Award type:** 2
- **Project period:** 1979-01-01 → 1997-11-30

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/3126290

## Citation

> US National Institutes of Health, RePORTER application 3126290, STREPTOCOCCUS PLASMIDS--MOLECULAR GENETIC ANALYSIS (2R01AI015619-14). Retrieved via MORVS 2026-10-25 from https://api.morvs.ai/grant/nih/3126290. Licensed CC0.

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