1
From the Department of Pathology, Queen’s University, Kingston, Ontario, Canada
,
Colleen Notley
1
From the Department of Pathology, Queen’s University, Kingston, Ontario, Canada
,
Shawn Tinlin
1
From the Department of Pathology, Queen’s University, Kingston, Ontario, Canada
,
Lisa Sawyer
1
From the Department of Pathology, Queen’s University, Kingston, Ontario, Canada
,
David Lillicrap
1
From the Department of Pathology, Queen’s University, Kingston, Ontario, Canada
› Author AffiliationsMany thanks to Dr. Robert Kisilevsky, John Anscin and Ruth Tan for help with the mouse model. We also thank Leslie Steele for helpful discussion and initial contributions to the development of this project. This project was funded by grants B-3123 from the Heart and Stroke Foundation of Ontario and MT-10912 from the Medical Research Council of Canada. M. B. is funded by a Studentship from the Medical Research Council of Canada. D. L. is a Career Investigator of the Heart and Stroke Foundation of Ontario.
Coagulation Factor VIII is an acute phase protein in humans that has recently been shown to be transcriptionally responsive to interleukin-6. In this study, we have demonstrated that the human Factor VIII promoter is activated in cultured hepatocytes exposed to bacterial lipopolysaccharide (LPS). Deletion analysis has narrowed the LPS-responsive element of the Factor VIII promoter to a small region which contains two C/EBP binding sites and an adjacent NFκB binding site. Mutation of the downstream C/EBP site reduces LPS-responsiveness by ∼50%, while mutation of the NFκB binding site completely eliminates LPS-responsiveness. While binding of C/EBPβ and NFκB is still observed in gel retardation studies using acute phase nuclear extracts and a probe containing mutations to the downstream C/EBP site, neither NFκB nor C/EBP appear to bind to a probe in which the NFκB site has been mutated. Conservation of this region of the Factor VIII promoter in species which exhibit an increase in Factor VIII levels in response to inflammatory stimuli suggests that these transcription factor binding sites are important for normal regulation of the Factor VIII gene under conditions of stress.
* Current Address: Dr. M. Begbie, National Heart and Lung Institute, Imperial College of Science and Technology and Medicine, Hammersmith Hospital Campus, Du Cane Road, London, W12 ONN, UK
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