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Isolation and Characterization of the cDNA for Mouse Neutrophil
Collagenase: Demonstration of Shared Negative Regulatory Pathways for
Neutrophil Secondary Granule Protein Gene Expression
Nathan D. Lawson,
Arati Khanna-Gupta, and
Nancy Berliner
From the Department of Biology and Section of Hematology, Department
of Internal Medicine, Yale University School of Medicine, New Haven,
CT.
A characteristic of normal neutrophil maturation is the induction of
secondary granule protein (SGP) mRNA expression. Several leukemic human
cell lines mimic normal morphologic neutrophil differentiation but fail
to express SGPs, such as lactoferrin (LF) and neutrophil gelatinase
(NG). In contrast, two murine cell lines (32D C13 and MPRO) are able to
differentiate into neutrophils and induce expression of LF and NG.
Therefore, to study the normal regulation and function of these genes,
the corresponding murine homologs must be isolated. Using cDNA
representational difference analysis (RDA) to compare a committed
myeloid progenitor cell line (EPRO) with the multipotent stem cell line
from which it was derived (EML), we isolated a fragment bearing
homology to human neutrophil collagenase (hNC). Here, we describe the
cloning and characterization of a full-length (~2 kb) clone that
exhibits nearly 65% nucleotide and 73% amino acid identity to hNC.
Ribonuclease protection analysis (RPA) of the tissues and cell lines
shows that mouse NC (mNC) is expressed only in cell lines exhibiting neutrophilic characteristics, further confirming its identity as the
mouse homolog of hNC. Furthermore, we have demonstrated a shared
negative regulatory pathway for this and other SGP genes. We have
previously shown that CCAAT displacement protein (CDP/cut) binds to a
specific region of the LF promoter, and overexpression of CDP blocks
G-CSF-induced upregulation of LF gene expression in 32D C13
cells. We show here that in these cells, upregulation of both
NC and NG is also blocked. CDP is thus the first identified transcription factor that is a candidate for mediating the shared regulation of neutrophil SGP protein genes.
Blood, Vol. 91 No. 7 (April 1), 1998:
pp. 2517-2524
© 1998 by The American Society of Hematology.

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