# GENETICS AND BIOCHEMISTRY OF INTRACELLULAR PROTEOLYSIS

> **NIH NIH R01** · CASE WESTERN RESERVE UNIVERSITY · 1987 · $169,217

## Abstract

The first major goal of this project is to complete the genetic analysis of
the physiological function of peptidases in Salmonella typhimurium.  To do
this we will:  a) isolate mutations affecting all of the remaining major
peptidases of the cell; b) use these mutants to define the role of the
individual enzymes in the protein turnover pathway and to study the effect
of their loss on cell growth and viability; and, c) characterize the
regulation of these peptidase genes.  Studies of peptidase regulation will
require analysis of DNA sequence changes and studies of the role of
individual peptidases on the degradation pathway will require purified
enzymes.  We therefore propose to isolate cloned pep genes for use in
obtaining DNA for the analysis of sequence changes in regulatory mutants
and for use in obtaining enzyme overproducers.  The second major goal of
the project is to begin genetic analysis of endoprotease function in
Salmonella.  We plan to isolate and genetically characterize mutations
affecting the major endoproteases of the cell.  We intend to use peptide
substrates in screening techniques similar to those already used for the
isolation of peptidase mutants.  Characterization of the physiological
consequences of these mutations should show which enzymes carry out which
steps in the protein degradation pathway and in other proteolytic
processes.  The sequence of steps by which a protein molecule is degraded
constitutes a major metabolic pathway occurring in all cells.  Little is
known about the roles of individual enzymes in this pathway in any
organism.  The use of a genetic approach to the study of this problem seems
especially appropriate given the large number of peptidases and proteases
and the broad overlap in their substrate specificities.  Well developed
techniques allow the construction of organisms lacking specific enzyme
activities or combinations of activities.  The usefulness of bacteria for
the production of peptide and protein gene products of cloned DNA from
non-bacterial sources probably depends at least in part on the construction
of strains that will not proteolytically destroy these products.  An
understanding of the specificities and regulation of cellular proteases and
peptidases may be crucial for the construction of such strains.

## Key facts

- **NIH application ID:** 3124708
- **Project number:** 5R01AI010333-16
- **Recipient organization:** CASE WESTERN RESERVE UNIVERSITY
- **SAM.gov UEI:** HJMKEF7EJW69
- **Principal Investigator:** CHARLES G MILLER
- **Activity code:** R01 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 1987
- **Award amount:** $169,217
- **Award type:** 5
- **Project period:** 1978-04-01 → 1988-03-31

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/3124708

## Citation

> US National Institutes of Health, RePORTER application 3124708, GENETICS AND BIOCHEMISTRY OF INTRACELLULAR PROTEOLYSIS (5R01AI010333-16). Retrieved via MORVS 2026-10-25 from https://api.morvs.ai/grant/nih/3124708. Licensed CC0.

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