Nucleotide Structure and Characterization of the Murine Gene Encoding the Endothelial Cell Protein C Receptor

Zhong Liang; Elliot D. Rosen; Francis J. Castellino

doi:10.1055/s-0037-1614529

Thrombosis and Haemostasis, Table of Contents

Thromb Haemost 1999; 81(04): 585-588
DOI: 10.1055/s-0037-1614529

Rapid Communication

Schattauer GmbH

Nucleotide Structure and Characterization of the Murine Gene Encoding the Endothelial Cell Protein C Receptor

Authors

Zhong Liang

¹From the Department of Chemistry and Biochemistry and the W. M. Keck Center for Transgene Research, University of Notre Dame, Notre Dame, Indiana, USA
Elliot D. Rosen

¹From the Department of Chemistry and Biochemistry and the W. M. Keck Center for Transgene Research, University of Notre Dame, Notre Dame, Indiana, USA
Francis J. Castellino

¹From the Department of Chemistry and Biochemistry and the W. M. Keck Center for Transgene Research, University of Notre Dame, Notre Dame, Indiana, USA

Abstract

Summary

The nucleotide sequence of the entire gene encoding the murine endothelial cell receptor for activated protein C (EPCR) has been determined. A total of 5303 bp of DNA was sequenced that included 4 exons and three introns, which constituted the coding region of the gene, as well as 393 bp upstream of the first exon and 841 bp downstream of the last exon. From the locations of the introns in this gene and analysis of the exon structures, it is clear the EPCR gene is a member of the CD1 class of multiple histocompatibility proteins, and its cDNA sequence is nearly identical to that of CCD41, a centrosome-associated protein. All elements needed for RNA polymerase II-based transcription are predicted to exist in the 5’ uncoded region of the gene, and potential 3’ regulatory sequences for efficient polyadenylation have been located at their optimal locations. A variety of highly probable transcription factor binding sites have been located in the 5’ region of the gene. These data suggest that the EPCR gene is under efficient transcriptional control, and support the finding that this gene product may be involved in the inflammatory pathway.

Full Text

References

References
1 Kisiel W, Canfield WM, Ericsson LH, Davie EW. Anticoagulant properties of bovine plasma protein C following activation by thrombin. Biochemistry 1977; 16: 5824-31.
2 Vehar GA, Davie EW. Preparation and properties of bovine factor VIII (antihemophilie factor). Biochemistry 1980; 19: 401-10.
3 Bajzar L, Morser J, Nesheim M. TAFI, or plasma procarboxypeptidase B, couples the coagulation and fibrinolytic cascades through the thrombinthrombomodulin complex. J Biol Chem 1996; 271: 16603-8.
4 Taylor FB, Lockhart MS. A new function for activated protein C: Activated protein C prevents inhibition of plasminogen activators by releasate from mononuclear leukocytes-platelet suspensions stimulated by phorbol diester. Thromb Res 1985; 37: 639-49.
5 Jalbert LR, Rosen ED, Moons L, Chan JYC, Carmeliet P, Collen D, Castellino FJ. Inactivation of the gene for anticoagulant protein C causes lethal perinatal consumptive coagulopathy in mice. J Clin Invest 1998 in press.
6 Taylor FB, Chang A, Esmon CT, D’Angelo A, Vigano-D’Angelo S, Blick KE. Protein C prevents the coagulopathic and lethal effects of Escherichia coli infusion in the baboon. J Clin Invest 1987; 79: 918-25.
7 Gerson WT, Dickerman JD, Bovill EG, Golden E. Severe acquired protein C deficiency in purpura fulminans associated with disseminated intravascular coagulation. Treatment with protein C concentrate. Pediatrics 1993; 91: 418-22.
8 Hogg PJ, Öhlin A-K, Stenflo J. Identification of structural domains in protein C involved in its interaction with thrombin-thrombomodulin on the surface of endothelial cells. J Biol Chem 1992; 267: 703-6.
9 Fukudome K, Esmon CT. Identification, cloning, and regulation of a novel endothelial cell protein C/activated protein C receptor. J Biol Chem 1994; 269: 26486-91.
10 Hancock WW, Grey ST, Hau L, Akalin E, Orthner C, Sayegh MH, Salem HH. Binding of activated protein C to a specific receptor on human mononuclear phagocytes inhibits intracellular calcium signaling and monocyte-dependent proliferative responses. Transplantation 1995; 60: 1525-32.
11 Benton WD, Davis RW. Screening λgt recombinant closs by hybridization to single plaques in situ. Science 1977; 196: 180-2.
12 Fukudome K, Esmon CT. Molecular cloning and expression of murine and bovine endothelial cell protein C/activated protein C receptor (EPCR). The structural and functional conservation in human, bovine, and murine EPCR. J Biol Chem 1995; 270: 5571-7.
13 Quandt K, Frech K, Karas H, Wingender E, Werner T. MatInd and MatInspector: new fast versatile tools for detection of consensus matches in nucleotide sequence data. Nucl Acids Res 1995; 23: 4878-84.
14 Calabi F, Jarvis JM, Martin L, Milstein C. Two classes of CD1 genes. Eur J Immunol 1989; 19: 285-92.
15 Ronne H, Widmark E, Rask L, Peterson PA. Intron sequences reveal evolutionary relationships among major histocompatibility complex class I genes. Proc Natl Acad Sci USA 1985; 82: 5860-4.
16 Calabi F, Belt KT, Yu CY, Bradbury A, Mandy WJ, Milstein C. The rabbit CD1 and the evolutionary conservation of the CD1 gene family. Immuno-genetics 1989; 30: 370-7.
17 Hughes AL. Evolutionary origin and diversification of the mammalian CD1 antigen genes. Mol Biol Evol 1991; 8: 185-201.
18 Rothbarth K, Petzelt C, Lu X, Todorov IT, Joswig G, Pepperkok R, Ansorge W, Werner D. cDNA-derived molecular characteristics and antibodies to a new centrosome-associated and G2/M phase-prevalent protein. J Cell Sci 1993; 104: 19-30.
19 Bucher P. Weight matrix descriptions of four eukaryotic RNA polymerase II elements derived from 502 unrelated promoter sequences. J Mol Biol 1990; 212: 563-78.
20 McLauchlan J, Gaffney D, Whitton JL, Clements JB. The consensus sequence YGTGTTYY located downstream from the AATAAA signal is required for efficient formation of mRNA 3U termini. Nucl Acids Res 1985; 13: 1347-68.
21 Akira S, Isshiki H, Sugita T, Tanabe O, Kinoshita S, Nishio Y, Nakajima T, Hirano T, Kishimoto T. A nuclear factor for IL-6 expression (NF-IL6) is a member of the C/EBP family. EMBO J 1990; 9: 1897-906.