# STRUCTURE-FUNCTION ANALYSIS OF SPECIFIC HUMAN GENES

> **NIH NIH R37** · BOSTON CHILDREN'S HOSPITAL · 1994 · $249,808

## Abstract

A goal of this laboratory's efforts is to employ molecular genetics
to understand the structure, regulation, and function of specific
human genes that are expressed in hematopoietic cells and
relevant to disease.  An important host defense system of
phagocytes is a plasma membrane associated NADPH oxidase that
reduces molecular oxygen to superoxide.  The components of the
oxidase system, their correspondence with specific genetic loci,
and the primary protein defect(s) in chronic granulomatous
disease (CGD) (a spectrum of genetic disorders that incapacitates
this enzyme system) have been largely unresolved.  Recently, the
gene and cDNA for the product lacking in X-linked CGD, the
classical inherited disease in the system, was cloned by "reverse
genetics" and shown to encode a 90 kd membrane glycoprotein
subunit of the phagocyte b-cytochrome.

In the proposed research the structure, expression, and function of
the 90 kd X-CGD protein will be pursued from a molecular
perspective.  Attention will be directed toward delineating the
cis-acting DNA elements that regulate lineage specific
(phagocytic) expression of the gene by gene transfer of
promoter/enhancer fusions and toward identification and
characterization of nuclear DNA-binding proteins that interact
with the critical DNA regions.  To study the X-CGD protein and
its function antibodies to specific domains will be generated to
determine its membrane orientation and to modulate the activity
of the system form the extracellular space.  A functional assay
for the protein will be established by transfer of full-length cDNA
into heterologous phagocytic and non-phagocytic cell lines, and
into deficient myelomonocytic cells.  With this assay and in vitro
mutagenesis of the cDNA the role of specific domains of the
protein and interaction with a 22 kd component of the phagocyte
b-cytochrome will be approached directly.  To study the
interaction of the 90 kd and 22 kd components, cDNA for the 22
kd will be isolated and used in coexpression experiments.  Finally,
the genetics of specific X-CGD mutations will be investigated,
especially in variant patients with residual b-cytochrome activity,
and RFLP probe suitable for improved prenatal diagnosis of X-
CGD by DNA analysis will be developed.

## Key facts

- **NIH application ID:** 2197707
- **Project number:** 5R37HD018661-12
- **Recipient organization:** BOSTON CHILDREN'S HOSPITAL
- **SAM.gov UEI:** Z1L9F1MM1RY3
- **Principal Investigator:** STUART H ORKIN
- **Activity code:** R37 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 1994
- **Award amount:** $249,808
- **Award type:** 5
- **Project period:** 1983-07-01 → 1998-06-30

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/2197707

## Citation

> US National Institutes of Health, RePORTER application 2197707, STRUCTURE-FUNCTION ANALYSIS OF SPECIFIC HUMAN GENES (5R37HD018661-12). Retrieved via MORVS 2026-10-25 from https://api.morvs.ai/grant/nih/2197707. Licensed CC0.

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