Domain swapping between FEN-1 and XPG defines regions in XPG that mediate nucleotide excision repair activity and substrate specificity
| Autor: | Marcel Hohl, Lidija Staresincic, Isabelle Dunand-Sauthier, Mauro Modesti, Stuart G. Clarkson, Pascale Jaquier-Gubler, Fabrizio Thorel, Orlando D. Schärer |
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| Přispěvatelé: | University of Zurich, Schärer, Orlando D |
| Rok vydání: | 2007 |
| Předmět: |
DNA Repair
Flap Endonucleases Ultraviolet Rays DNA repair DNA damage Recombinant Fusion Proteins Flap structure-specific endonuclease 1 Biology DNA-binding protein Substrate Specificity 03 medical and health sciences 0302 clinical medicine 1311 Genetics Genetics Humans Flap endonuclease Cells Cultured 030304 developmental biology 0303 health sciences Nuclease Nucleic Acid Enzymes 10061 Institute of Molecular Cancer Research DNA replication Nuclear Proteins Endonucleases Molecular biology Protein Structure Tertiary Cell biology DNA-Binding Proteins 030220 oncology & carcinogenesis biology.protein 570 Life sciences biology DNA Damage Transcription Factors Nucleotide excision repair |
| Zdroj: | Nucleic Acids Research |
| ISSN: | 1362-4962 0305-1048 |
| DOI: | 10.1093/nar/gkm092 |
| Popis: | FEN-1 and XPG are members of the FEN-1 family of structure-specific nucleases, which share a conserved active site. FEN-1 plays a central role in DNA replication, whereas XPG is involved in nucleotide excision repair (NER). Both FEN-1 and XPG are active on flap structures, but only XPG cleaves bubble substrates. The spacer region of XPG is dispensable for nuclease activity on flap substrates but is required for NER activity and for efficient processing of bubble substrates. Here, we inserted the spacer region of XPG between the nuclease domains of FEN-1 to test whether this domain would be sufficient to confer XPG-like substrate specificity and NER activity on a related nuclease. The resulting FEN-1-XPG hybrid protein is active on flap and, albeit at low levels, on bubble substrates. Like FEN-1, the activity of FEN-1-XPG was stimulated by a double-flap substrate containing a 1-nt 3′ flap, whereas XPG does not show this substrate preference. Although no NER activity was detected in vitro, the FEN-1-XPG hybrid displays substantial NER activity in vivo. Hence, insertion of the XPG spacer region into FEN-1 results in a hybrid protein with biochemical properties reminiscent of both nucleases, including partial NER activity. |
| Databáze: | OpenAIRE |
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