# STUDIES OF EPIDERMAL MACROMOLECULAR METABOLISM

> **NIH NIH R01** · NEW YORK UNIVERSITY · 1991 · $213,288

## Abstract

Our long-term goal is to elucidate the pathways and controls of epidermal
keratin synthesis in health and disease.  During the next project period we
shall focus on the control of expression of genes for the six major
epidermal keratins:  K14 and K5 of basal cells, K10 and K1 of suprabasal
differentiated cells and K16 and K6 of hyperproliferative epidermis.  Our
specific aims are to characterize the cis-active regulatory regions of the
keratin genes important for transcriptional control, to define the
transacting factors which interact with their regulatory regions, to
evaluate the changes in transcriptional control associated with
hyperproliferative diseases such as psoriasis and to determine the
interactions among the regulatory elements involved in coordinate
expression of keratin gene expression in vivo.

Initially, we will identify the cis- and trans-acting factors which
influence normal keratin genes.  The gene for keratin K14, the acidic
member of the basal cell-specific pair, will serve as a paradigm for our
subsequent studies of the genes coding for the other keratins of normal and
pathologic epidermis.  The regulatory regions (promoters, enhancers and
negative regulatory elements) will be identified by transfection into cells
of constructs which contain the regulatory regions of the keratin genes
linked to the chloramphenicol-acetyl-transferase (CAT) reporter gene.
Sequences identified as involved in control will be amplified by the
polymerase chain reaction and studied in more detail in CAT assays.

Trans-acting factors will be studied in 4 cell lines which vary in their
expression of the keratins under study (fibroblasts - none; HeLa - K18 and
K8; SCC 12 - K14, K16, K5, K6; NM1 - K14, K16, K10, K5, K6, K1).  The
factors will be identified by gel retardation, footprinting and methylation
interference assays.  Mutations produced in the cis-active regulatory
regions (through the polymerase chain reaction) will prove the functional
significance of trans-acting factors.  Parallel studies will then be done
with cultured human epidermal cells, normal and psoriatic epidermis so that
we begin to document the changes in transcriptional control associated with
disease.

Finally, we will study the interactions among the regulatory factors for
the six human epidermal keratin genes.  The experiments will require co-
transfection studies and ultimately transgenic mice.  Our goal will be to
understand the coordinate expression which is so important in this large,
clinically significant gene family.

## Key facts

- **NIH application ID:** 3155869
- **Project number:** 5R01AR030682-10
- **Recipient organization:** NEW YORK UNIVERSITY
- **SAM.gov UEI:** NX9PXMKW5KW8
- **Principal Investigator:** IRWIN M FREEDBERG
- **Activity code:** R01 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 1991
- **Award amount:** $213,288
- **Award type:** 5
- **Project period:** 1981-07-01 → 1994-06-30

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/3155869

## Citation

> US National Institutes of Health, RePORTER application 3155869, STUDIES OF EPIDERMAL MACROMOLECULAR METABOLISM (5R01AR030682-10). Retrieved via MORVS 2026-10-25 from https://api.morvs.ai/grant/nih/3155869. Licensed CC0.

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