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Thromb Haemost 2009; 102(03): 469-478
DOI: 10.1160/TH09-03-0162
Blood Coagulation, Fibrinolysis and Cellular Haemostasis
Schattauer GmbH

The recruitment of blood coagulation factor X into snake venom gland as a toxin

The role of promoter Cis-elements in its expression
Shiyang Kwong
1   Department of Biological Sciences, Faculty of Science, National University of Singapore, Singapore, Singapore
,
Anthony E. Woods
2   Sansom Institute, School of Pharmacy and Medical Sciences, University of South Australia, Adelaide, South Australia, Australia
,
Peter J. Mirtschin
2   Sansom Institute, School of Pharmacy and Medical Sciences, University of South Australia, Adelaide, South Australia, Australia
3   Venom Supplies Pty Ltd, Tanunda, South Australia, Australia
,
Ruowen Ge
1   Department of Biological Sciences, Faculty of Science, National University of Singapore, Singapore, Singapore
,
R. Manjunatha Kini
1   Department of Biological Sciences, Faculty of Science, National University of Singapore, Singapore, Singapore
4   Department of Biochemistry, VCU Medical Center, Virginia Commonwealth University, Richmond, Virginia, USA
› Author AffiliationsFinancial support: This work is supported by the Academic Research Grants from the National University of Singapore.
Further Information

Publication History

Received: 12 March 2009

Accepted after minor revision: 14 May 2009

Publication Date:
22 November 2017 (online)

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Summary

Trocarin D is a prothrombin activator from the Tropidechis carinatus venom. It is a functional and structural homologue to mammalian blood coagulation factor Xa.Trocarin D is hypothesised to have evolved from its factor X counterpart (TrFX) through gene duplication and recruitment.The genes of trocarin D and TrFX have significant sequence identities, except for insertions/deletions in their intron 1 and promoter regions. In trocarin D intron 1 region, there are three insertions and two deletions. In trocarin D promoter region, there is a novel 264 bp insertion which has potential cis-elements.This insertion is termed as Venom Recruitment/Switch Element (VERSE) and is hypothesised to account for switching the low-level constitutive expression of factor X in the liver to the high-level inducible expression of trocarin D in the venom gland. To understand the role of VERSE in the trocarin D expression,its cis-elements were characterised by luciferase assays in mammalian cell lines as well as snake venom gland cells. The ability of VERSE to drive luciferase expression is comparable to that of the trocarin D promoter. The predicted cis-elements are important in promoting expression as their mutagenesis resulted in lower luciferase expression.VERSE minimal core promoter and three novel cis-elements (two up-regulatory and one suppressor elements) were identified using deletion/site-directed mutagenesis studies. VERSE is primarily responsible for the increase of trocarin D expression. The insertions/deletions within trocarin D intron 1 need to be characterised for their role in tissue-specific and inducible expression of trocarin D.

Keywords

Factor X - promoter - prothrombin activator - recruitment - snake venom
 

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