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Cloning and characterization of human complement component C7 promoter

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Abstract

To study the transcriptional regulation of the human complement component C7, a 1 kb promoter fragment was cloned and the transcription start site was determined. C7 is expressed by the hepatoma-derived cell line Hep-3B, but not by Hep-G2. Transfection of these cell lines with different C7 promoter–luciferase constructs demonstrated that 1 kb of the 5′-flanking region contains the necessary elements for driving C7 transcription in a tissue-specific manner and showed that the sequence between −29/+102 retained the majority of C7 promoter activity in Hep-3B. Electrophoretic mobility shift assays suggested that the binding of the C/EBPα transcription factor to a C/EBP sequence located at +42 is essential for C7 expression. To investigate whether the absence of C/EBPα expression in Hep-G2 cells is responsible for the lack of C7 transcription, Hep-G2 cells were transfected with a C/EBPα expression vector. C/EBPα transactivated the C7 luciferase reported gene and restored the C7 expression in Hep-G2 cells.

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Acknowledgements

We thank Dr R Wuzner for providing us with the anti-C7 antibodies, Dr M Hobart for providing us with the YAC clones, Dr GJ Darlington for the kind gift of the C/EBPα expression vector and David H Wallace (Member of the Council of Biology Editors and the Association of European Science Editors) for critical revision of the manuscript. This work was supported in part by a Spanish Ministry of Health grant Fis 00/0208.

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Authors and Affiliations

  1. Functional Biology Department, University of Oviedo, Spain

    S González

  2. Department of Immunology, Hospital Central de Asturias, Oviedo, Spain

    J Martínez-Borra & C López-Larrea

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  1. S González
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  2. J Martínez-Borra
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  3. C López-Larrea
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Correspondence to C López-Larrea.

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González, S., Martínez-Borra, J. & López-Larrea, C. Cloning and characterization of human complement component C7 promoter. Genes Immun 4, 54–59 (2003). https://doi.org/10.1038/sj.gene.6363902

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