Expression of the Drosophila melanogaster S3 ribosomal/repair protein in T24 human bladder cells

Monica Ropolo, Alessandro Geroldi, Ottavio Rossi, Paolo Degan, Simona Zupo, Alessandro Poggi, Guido Frosina

Research output: Contribution to journalArticlepeer-review

Abstract

Repair of 8-oxo-7,8-dihydroguanine (8-oxoG) is inefficient in human cells due to the poor catalytic properties of hOGG1, the major DNA glycosylase involved in the removal of this oxidized base. The S3 ribosomal/repair protein from Drosophila melanogaster (dS3) is endowed with a potent 8-oxoG glycolytic activity coupled with α β, δ-AP lyase. In vitro repair experiments have shown that pure GST-tagged dS3 can stimulate a >40-fold increase in the rate of 8-oxoG repair by human cell extracts. In this study, we expressed dS3 fused to the Enhanced Green Fluorescent Protein (EGFP) in T24 human bladder cells in order to accelerate the repair of 8-oxoG in vivo. Limiting dilution and Fluorescence-Activated Cell Sorting (FACS) were used in an effort to isolate cells with elevated EGFP-dS3 expression; however, the cells that were isolated invariably had severe growth impairment. Curiously, EGFP-dS3 expression was slightly increased after recovering cells from liquid nitrogen, but it was not possible under those conditions to achieve a significant acceleration of 8-oxoG repair. The data confirm and extend our previous results obtained with Chinese hamster CHO cells and indicate that elevated expression of dS3 may be toxic to at least some types of mammalian cells, thus limiting its use in vivo as a protective factor against oxidative DNA damage.

Original languageEnglish
Pages (from-to)3811-3818
Number of pages8
JournalAnticancer Research
Volume24
Issue number6
Publication statusPublished - Nov 2004

Keywords

  • Bladder cells
  • Drosophila melanogaster S3
  • dS3
  • Oxidative DNA damage
  • Ribosomal/repair protein

ASJC Scopus subject areas

  • Cancer Research
  • Oncology

Fingerprint Dive into the research topics of 'Expression of the Drosophila melanogaster S3 ribosomal/repair protein in T24 human bladder cells'. Together they form a unique fingerprint.

Cite this