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(Received for publication, January 17, 1996) In an attempt to correlate the cell surface expression of
Le
Volume 271,
Number 17,
Issue of April 26, 1996 pp. 10317-10328
©1996 by The American Society for Biochemistry and Molecular Biology, Inc.
1,3-L
-Fucosyltransferase
Expression in Developing Human Myeloid Cells
ANTIGENIC, ENZYMATIC, AND mRNA ANALYSES
and sialyl-Le
structures in immature and
mature myeloid cells with the genes expressing
1,3-fucosyltransferase(s) we have examined: 1) the properties of
the cellular
1,3-fucosyltransferases and the mRNA transcripts
corresponding to the five cloned genes, Fuc-TIII, Fuc-TIV, Fuc-TV,
Fuc-TVI, and Fuc-TVII, in mature granulocytes and in the myeloid cell
line HL-60, before and after dimethyl sulfoxide-induced differentiation
and 2) the properties of the
1,3-fucosyltransferases expressed in
COS-7 cells transfected with plasmids containing Fuc-TIV and Fuc-TVII
cDNAs. The previously shown increase in cell surface expression of
sialyl-Le
on differentiation of HL-60 cells (Skacel P. O.,
Edwards A. J., Harrison C. T., and Watkins W. M.(1991) Blood 78, 1452-1460) is accompanied by a sharp fall in expression
of Fuc-TIV mRNA and a persistence of expression of Fuc-TVII mRNA. The
properties of the
1,3-fucosyltransferase expressed in COS-7 cells
transfected with Fuc-TIV are consistent with this being the major gene
responsible for the expression of Le
in the immature
myeloid cells. In Northern blot analyses, no transcripts of Fuc-TIII,
Fuc-TV, or Fuc-TVI were detected in total RNA from mature granulocytes
or mRNA from HL-60 cells before or after differentiation. In total RNA
from mature granulocytes, Fuc-TIV transcripts were only faintly
visible, whereas Fuc-TVII transcripts were quite definitely expressed.
The specificity properties of Fuc-TVII expressed in COS-7 cells are
consistent with this gene being the major candidate
1,3-fucosyltransferase controlling the expression of
sialyl-Le
on mature cells. However, Le
continues to be expressed on the surface of mature granulocytes and
cell extracts retain the capacity to transfer fucose to non-sialylated
acceptor substrates. The question therefore remains as to whether these
properties result from the weakly expressed Fuc-TIV gene or whether
another
1,3-fucosyltransferase gene remains to be identified.
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