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Isolation of a novel macrophage-specific gene by differential cDNA analysis
K Spilsbury, MA O'Mara, WM Wu, PB Rowe, G Symonds and Y Takayama
Children's Medical Research Institute, Westmead, Australia.
To analyze myelomonocytic differentiation we have used the approach of
differential cDNA analysis to isolate novel genes that are preferentially
expressed in mature macrophages. Differential screening of a macrophage
cDNA library led to the identification of a novel cDNA that showed
macrophage lineage- and differentiation stage-specific expression.
Transcripts from the gene, which we have termed Mpg-1, are found at a high
level in mature human and murine macrophages and at a moderate level in
certain myelomonocytic cell lines. The expression of Mpg-1 was found to
increase when murine fetal liver hematopoietic progenitor cells were
induced to differentiate into macrophages. An Mpg- 1-specific transcript
was not detected in a wide variety of other tissues and cell lines. The DNA
sequence of Mpg-1 (4,214 bp) was obtained from a series of overlapping
cDNA, 3' rapid amplification of cDNA ends (RACE), and genomic clones.
Primer extension analysis predicted the existence of multiple transcription
start sites, ranging from 26 to 117 bp upstream of the 5' proximal ATG of
the open reading frame. The predicted 669-amino acid, Mpg-1-encoded protein
has potential glycosylation and phosphorylation sites in addition to a
signal sequence. The core protein is predicted to have a molecular weight
of 71 to 74 kD. Computer-assisted local similarity searches indicate that
Mpg-1 is a novel gene that may share a distant ancestry to perforin, a
lytic protein found in cytotoxic T lymphocytes and natural killer cells.
Volume 85,
Issue 6,
pp. 1620-1629,
03/15/1995
Copyright © 1995 by The American Society of Hematology
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