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© The Rockefeller University Press,
0021-9525/1999//563 $5.00
The Journal of Cell Biology, Volume 145, Number 3,
, 1999 563-577
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Mouse Ten-m/Odz Is a New Family of Dimeric Type II Transmembrane Proteins Expressed in Many Tissues
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
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.
Key Words: ten-m/odz • transmembrane protein • pair rule • epidermal growth factor
Abbreviations used in this paper: AP, alkaline phosphatase; CNS, central nervous system.
T. Oohashi was supported by the Japanese Society for Promotion of Science and by a Max Planck fellowship. X.-H. Zhou is a Wenner Gren fellow. This work was supported by the Swedish Medical Research Council and the Kanae Foundation for Life and Socio-medical Science.
The online version of this article contains supplemental material. T. Oohashi and X.-H. Zhou contributed equally to this work.
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