Thromb Haemost 1992; 67(02): 272-276
DOI: 10.1055/s-0038-1648424
Original Articles
Schattauer GmbH Stuttgart

Alternative Splicing Is Responsible for the Presence of Two Tissue Factor mRNA Species in LPS Stimulated Human Monocytes

C Paul
The Haemostasis and Thrombosis Research Unit, University Hospital, Leiden, The Netherlands
,
E van der Logt
The Haemostasis and Thrombosis Research Unit, University Hospital, Leiden, The Netherlands
,
Pieter H Reitsma
The Haemostasis and Thrombosis Research Unit, University Hospital, Leiden, The Netherlands
,
Rogier M Bertina
The Haemostasis and Thrombosis Research Unit, University Hospital, Leiden, The Netherlands
› Author Affiliations
Further Information

Publication History

Received 02 July 1991

Accepted after revision 05 August 1991

Publication Date:
02 July 2018 (online)

Summary

Although normally absent from the surface of all circulating cell types, tissue factor (TF) can be induced to appear on circulating monocytes by stimulants like bacterial lipopolysaccharide (LPS) and phorbolesters. Northern analysis of RNA isolated from LPS stimulated human monocytes demonstrates the presence of 2.2 kb and 3.1 kb TF mRNA species. The 2.2 kb message codes for the TF protein. As demonstrated by Northern blot analysis with a variety of TF gene probes, the 3.1 kb message arises from an alternative splicing process which fails to remove 955 bp from intron 1. Because of a stop codon in intron 1 no TF protein is produced from the 3.1 kb transcript. This larger transcript should therefore not be taken into account when comparing TF gene transcription and TF protein levels.

 
  • References

  • 1 Furie B, Furie BC. The molecular basis of blood coagulation. Cell 1988; 53: 505-518
  • 2 Rickies FR, Rick PR. Structural features of Salmonella typhimurium lipopolysaccharide required for activation of tissue factor in human mononuclear cells. J Clin Invest 1977; 59: 1188-1198
  • 3 Rothberger H, Zimmerman TS, Spiegelberg HL, Vaughan JH. Leukocyte procoagulant activity; Enhancement of production in vitro by IgG and Ag:Ab complexes. J Clin Invest 1977; 59: 549-557
  • 4 Edwards RL, Rickies FR. The role of Human T cells (and T cell products) for monocyte tissue factor generation. J Immunol 1980; 125: 606-609
  • 5 Lyberg T, Prydz H. Phorbolesters induce synthesis of thromboplastin activity in human monocytes. Biochem J 1981; 194: 699-706
  • 6 Gregory SA, Morrissey JH, Edgington TS. Regulation of tissue factor gene expression in the monocyte procoagulant response to endotoxin. Mol Cell Biol 1989; 9: 2752-2755
  • 7 Scarpati EM, Wen D, Broze GJ, Miletich JP, Flandermeyer RR, Siegel NR, Sadler IE. Human tissue factor: cDNA sequence and chromosome localization of the gene. Biochemistry 1987; 26: 5234-5238
  • 8 Mackman N, Morrissey JH, Fowler B, Edgington TS. Complete sequence of the human tissue factor gene, a highly regulated cellular receptor that initiates the coagulation protease cascade. Biochemistry 1989; 28: 1755-1762
  • 9 Angel P, Imagawa M, Chiu R, Stein B, Imbra I, Rahmsdorf HI, Jonat C, Herrlich P, Karin M. Phorbolester inducible genes contain a common Cis element recognized by a TPA modulated trans-acting factor. Cell 1987; 49: 729-739
  • 10 Imagawa M, Chiu R, Karin M. Transcription factor AP-2 mediates induction by two different signal-transduction pathways: Protein Kinase C and cAMP. Cell 1987; 51: 251-260
  • 11 Spicer EK, Horton R, Bloem L, Bach R, Williams KR, Guha A, Krans J, Lin TC, Nemerson Y, Königsberg WH. Isolation of cDNA clones coding for human tissue factor: Primary structure of the protein and cDNA. Proc Natl Acad Sci USA 1987; 84: 5148-5152
  • 12 Maniatis T, Fritsch EF, Sambrook J. Molecular Cloning: A Laboratory Manual. Cold Spring Harbor Laboratory; Cold Spring Harbor, NY: 1982
  • 13 Sanger F, Nicklen S, Coulson AR. DNA sequencing with chainterminating inhibitors. Proc Natl Acad Sci USA 1977; 74: 5463-5467
  • 14 Chomczymski P, Sacchi N. Single step method of RNA acid guanidium thiocyanate-phenol-chloroform extraction. Anal Biochem 1987; 162: 156-159
  • 15 Breathnach R, Chambón P. Organization and expression of eucary-otic split genes coding for proteins. Annu Rev Biochem 1981; 50: 349-383
  • 16 Caput D, Bender B, Hartog K, Thayer R, Brown-Shimer S, Cerami A. Identification of a common nucleotide sequence in the 3’untrans-lated region of mRNA molecules specifying inflammatory mediators. Proc Natl Acad Sci USA 1986; 83: 1670-1674
  • 17 Shaw G, Kamen R. A conserved AU sequence from the 3’untrans-lated region of GM-CSF mRNA mediates selective mRNA degradation. Cell 1986; 46: 659-667
  • 18 Gidoni D, Dynan WS, Tjian R. Multiple specific contacts between a mammalian transcription factor and its cognate promoters. Nature 1984; 321: 409-413