Inhibition of activating transcription factor 1- and cAMP-responsive element-binding protein-activated transcription by an intracellular single chain Fv fragment.

The Journal of Biological Chemistry
J M BosilevacS H Hinrichs

Abstract

Activating transcription factor 1 (ATF1) and cAMP-responsive element (CRE)-binding protein (CREB) activate transcription through CREs located in the promoters of cellular and viral genes. We previously described a monoclonal antibody (mAb41.4) that prevents ATF1 binding to DNA and reduces CRE-driven promoter activity in vitro (Orten, D. J., Strawhecker, J. M., Sanderson, S. D., Huang, D., Prytowsky, M. B. , and Hinrichs, S. H. (1994) J. Biol. Chem. 269, 32254-32263). A single chain Fv (scFv) fragment from the mAb41.4-expressing hybridoma was generated to provide a means to investigate transcription factor function via intracellular expression of the scFv fragment. The affinity of scFv4 (subgroup: VL kappa-III, VH miscellaneous) for ATF1 was similar to that of the parental mAb and the Fab fragment, but it demonstrated greater inhibitory activity and reacted with CREB. scFv4 disrupted the binding of both ATF1 and CREB in electrophoretic mobility shift assays and reduced expression of CRE-driven expression in vitro. Transient expression of scFv had no effect on the non-CRE-containing adenovirus major late promoter. The proliferating cell nuclear antigen promoter, containing two CREs, was significantly more sensitive to inhibition ...Continue Reading

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Citations

May 27, 1999·Journal of Biomedical Science·A Krause BoehmS H Hinrichs
Jan 22, 2003·Biochemical and Biophysical Research Communications·Olga EstebanMariano Cambra
Jun 4, 2008·Sarcoma·R J OlsenS H Hinrichs
Feb 19, 2000·Biochemical and Biophysical Research Communications·M K SongI H Choi
Jun 20, 2006·Microbes and Infection·Christer LarssonSven Bergström
Nov 27, 1999·The Journal of Biological Chemistry·J M BosilevacS H Hinrichs

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