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J. Cell Biol.,
Volume 145, Number 3, May 3, 1999 563-577
* Max Planck Institute for Biochemistry, 82152 Martinsried, Germany; The Drosophila gene ten-m/odz is the only
pair rule gene identified to date which is not a transcription factor. In an attempt to analyze the structure and
the function of ten-m/odz in mouse, we isolated four
murine ten-m cDNAs which code for proteins of 2,700-2,800 amino acids. All four proteins (Ten-m1-4)
lack signal peptides at the NH2 terminus, but contain a
short hydrophobic domain characteristic of transmembrane proteins, 300-400 amino acids after the NH2 terminus. About 200 amino acids COOH-terminal to this
hydrophobic region are eight consecutive EGF-like domains.
Cell transfection, biochemical, and electronmicroscopic studies suggest that Ten-m1 is a dimeric type II
transmembrane protein. Expression of fusion proteins
composed of the NH2-terminal and hydrophobic domain of ten-m1 attached to the alkaline phosphatase reporter gene resulted in membrane-associated staining
of the alkaline phosphatase. Electronmicroscopic and
electrophoretic analysis of a secreted form of the extracellular domain of Ten-m1 showed that Ten-m1 is a disulfide-linked dimer and that the dimerization is mediated by EGF-like modules 2 and 5 which contain an
odd number of cysteines.
Northern blot and immunohistochemical analyses
revealed widespread expression of mouse ten-m genes,
with most prominent expression in brain. All four ten-m
genes can be expressed in variously spliced mRNA isoforms. The extracellular domain of Ten-m1 fused to an alkaline phosphatase reporter bound to specific regions
in many tissues which were partially overlapping with
the Ten-m1 immunostaining. Far Western assays and
electronmicroscopy demonstrated that Ten-m1 can
bind to itself.
Department of Molecular Biology and Biochemistry,
Okayama University, Medical School, Okayama 700, Japan; § Department of Experimental Pathology and Department of
Ophthalmology, Lund University, S-221 85 Lund, Sweden; and ¶ Friedrich Miescher Institut, 4058 Basel, Switzerland
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