PURPOSE OF INVESTIGATION: Regulatory elements of the 5'-flanking region of the DNA-repair gene Rad51 were analysed to characterise pathological alterations of Rad51 mRNA expression during tumour development. METHODS: Various fragments of the Rad51 promoter were cloned into the pGL3 reporter vector and the respective promoter activity was determined by luciferase assays in transfected U2-OS cells. Transcription factor binding was identified using Protein/DNA arrays. RESULTS: The region encompassing base pairs -204 to -58 was identified as crucial for Rad51 gene transcription. Down regulator sequences are present upstream (-305 to -204) and downstream (-48 and +204) of this core promoter element. Promoter activity is significantly enhanced by substituting G at the polymorphic positions +135 and +172 for C and T, respectively. Transcription factors Ets1/PEA3, E2F1, p53, EGR1, and Stat5 were identified as relevant for regulating expression of Rad51. CONCLUSION: We identified three separate cis-sequence elements within the Rad51 transcriptional promoter, one ensuring basal levels of expression and two elements limiting expression to relatively low levels. The characterisation of transcription factor binding might help to explain high-level expression of Rad51 in a variety of solid tumours. The polymorphic sites appear important for the increased risk of breast and/or ovarian cancer for BRCA2 mutation carriers.
PURPOSE OF INVESTIGATION: Regulatory elements of the 5'-flanking region of the DNA-repair gene Rad51 were analysed to characterise pathological alterations of Rad51 mRNA expression during tumour development. METHODS: Various fragments of the Rad51 promoter were cloned into the pGL3 reporter vector and the respective promoter activity was determined by luciferase assays in transfected U2-OS cells. Transcription factor binding was identified using Protein/DNA arrays. RESULTS: The region encompassing base pairs -204 to -58 was identified as crucial for Rad51 gene transcription. Down regulator sequences are present upstream (-305 to -204) and downstream (-48 and +204) of this core promoter element. Promoter activity is significantly enhanced by substituting G at the polymorphic positions +135 and +172 for C and T, respectively. Transcription factors Ets1/PEA3, E2F1, p53, EGR1, and Stat5 were identified as relevant for regulating expression of Rad51. CONCLUSION: We identified three separate cis-sequence elements within the Rad51 transcriptional promoter, one ensuring basal levels of expression and two elements limiting expression to relatively low levels. The characterisation of transcription factor binding might help to explain high-level expression of Rad51 in a variety of solid tumours. The polymorphic sites appear important for the increased risk of breast and/or ovarian cancer for BRCA2 mutation carriers.
Authors: Antonis C Antoniou; Olga M Sinilnikova; Jacques Simard; Mélanie Léoné; Martine Dumont; Susan L Neuhausen; Jeffery P Struewing; Dominique Stoppa-Lyonnet; Laure Barjhoux; David J Hughes; Isabelle Coupier; Muriel Belotti; Christine Lasset; Valérie Bonadona; Yves-Jean Bignon; Timothy R Rebbeck; Theresa Wagner; Henry T Lynch; Susan M Domchek; Katherine L Nathanson; Judy E Garber; Jeffrey Weitzel; Steven A Narod; Gail Tomlinson; Olufunmilayo I Olopade; Andrew Godwin; Claudine Isaacs; Anna Jakubowska; Jan Lubinski; Jacek Gronwald; Bohdan Górski; Tomasz Byrski; Tomasz Huzarski; Susan Peock; Margaret Cook; Caroline Baynes; Alexandra Murray; Mark Rogers; Peter A Daly; Huw Dorkins; Rita K Schmutzler; Beatrix Versmold; Christoph Engel; Alfons Meindl; Norbert Arnold; Dieter Niederacher; Helmut Deissler; Amanda B Spurdle; Xiaoqing Chen; Nicola Waddell; Nicole Cloonan; Tomas Kirchhoff; Kenneth Offit; Eitan Friedman; Bella Kaufmann; Yael Laitman; Gilli Galore; Gad Rennert; Flavio Lejbkowicz; Leon Raskin; Irene L Andrulis; Eduard Ilyushik; Hilmi Ozcelik; Peter Devilee; Maaike P G Vreeswijk; Mark H Greene; Sheila A Prindiville; Ana Osorio; Javier Benitez; Michal Zikan; Csilla I Szabo; Outi Kilpivaara; Heli Nevanlinna; Ute Hamann; Francine Durocher; Adalgeir Arason; Fergus J Couch; Douglas F Easton; Georgia Chenevix-Trench Journal: Am J Hum Genet Date: 2007-10-16 Impact factor: 11.025
Authors: S Franceschi; S Tomei; C M Mazzanti; F Lessi; P Aretini; M La Ferla; V De Gregorio; F Pasqualetti; K Zavaglia; G Bevilacqua; A G Naccarato Journal: J Neurooncol Date: 2015-10-28 Impact factor: 4.130
Authors: Anita Mitra; Charles Jameson; Yolanda Barbachano; Lydia Sanchez; Zsofia Kote-Jarai; Susan Peock; Nayanta Sodha; Elizabeth Bancroft; Anne Fletcher; Colin Cooper; Douglas Easton; Rosalind Eeles; Christopher S Foster Journal: Histopathology Date: 2009-12 Impact factor: 5.087