# SURFACE OLIGOSACCHARIDES IN EMBRYONAL CARCINOMA CELLS

> **NIH NIH R01** · UNIVERSITY OF GEORGIA · 1992 · $149,241

## Abstract

The major objectives of this proposal are to gain fundamental understanding
about the structure and steps in the biosynthesis and targeting of resident
proteins in the Golgi apparatus.  The work will focus on a specific
recombinant glycosyltransferase, namely the murine UDPGal:beta-D-Gal
alpha1,3 Galactosyltransferase (alpha1,3-GT).  There is currently little
information available about the biosynthesis of glycosyltransferases and
the factors regulating their intracellular expression.

Work in the previous funding period resulted in the cloning and sequencing
of the cDNA for this enzyme from the mouse teratocarcinoma cell line F9.
Both transient and permanent expression of the gene was accomplished in
heterologous cell types, including COS-1 cells and Chinese hamster ovary
(CHO) cells.  In the proposed work the structure and biosynthesis of the
enzyme in three different cell lines - F9 cells; F9 cells induced to
permanently differentiate with retinoic acid (RA/F9 cells), a treatment
causing elevated expression of both alpha1,3-GT transcripts and enzyme
activity; and COS-1 cells transfected with cDNA for alpha1,3-GT - will be
investigated.  The specific aims of this renewal application are to:

1. Determine the sites on the alpha1,3-GT for addition of Asn- and/or
Ser/Thr-linked oligosaccharides, structurally characterize the
oligosaccharides, and identify other potential posttranslational
modifications;

2. Define the biosynthesis of the alpha1,3-GT by following it from its
formation in the endoplasmic reticulum through the Golgi apparatus and to
the cell exterior, where it appears to be eventually secreted;

3. Explore the possibility that there are structural elements within the
alpha1,3-GT which target it to specific compartments of the Golgi apparatus
using specific mutagenesis procedures and expression in COS-1 cells; and

4. Define the factors responsible for proteolysis and release of the enzyme
from cells using the three cell lines mentioned above.

Successful completion of the studies proposed will provide fundamental
knowledge about the structure, biosynthesis, turnover, and intracellular
expression of a developmentally-regulated glycosyltransferase in animal
cells.  This work will also provide new information about the biosynthesis
of the Golgi apparatus and its components.

## Key facts

- **NIH application ID:** 3175421
- **Project number:** 5R01CA037626-08
- **Recipient organization:** UNIVERSITY OF GEORGIA
- **SAM.gov UEI:** NMJHD63STRC5
- **Principal Investigator:** RICHARD D CUMMINGS
- **Activity code:** R01 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 1992
- **Award amount:** $149,241
- **Award type:** 5
- **Project period:** 1988-07-01 → 1992-06-30

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/3175421

## Citation

> US National Institutes of Health, RePORTER application 3175421, SURFACE OLIGOSACCHARIDES IN EMBRYONAL CARCINOMA CELLS (5R01CA037626-08). Retrieved via MORVS 2026-10-25 from https://api.morvs.ai/grant/nih/3175421. Licensed CC0.

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