Differences in DNA-binding efficiency of Sp1 to aldolase and pyruvate kinase promoter correlate with altered redox states in resting and proliferating rat thymocytes.
Thymocytes induce their glycolytic enzymes as they undergo transition from the resting to the proliferating state. Corresponding increases in mRNA levels point to a transcriptional regulation. Electrophoretic mobility shift assays revealed that the DNA-binding efficiency of Sp1 is increased when nuclear extracts from proliferating compared to resting rat thymocytes were used. Here we demonstrate that hydrogen peroxide, added to nuclear extract from proliferating cells, decreases the Sp1 DNA-binding activity, whereas in nuclear extracts from resting cells dithioerythritol fully restores DNA-binding efficiency. Moreover we show that in contrast to resting thymocytes, production of reactive peroxide anions upon priming with phorbol 12-myristate 13-acetate is nearly abolished in the proliferating cells. From these results we propose that reactive oxygen intermediates affect the interaction of the Sp1 transcription factor with its consensus sequence and subsequently regulate glycolytic gene expression.
Schäfer
D
D
Institut für Biochemie, Medizinische Fakultät, Universität Erlangen-Nürnberg, Germany.
Hamm-Künzelmann
B
B
Hermfisse
U
U
Brand
K
K
D000818:Animals; D001665:Binding Sites; D002478:Cells, Cultured; D003208:Concanavalin A; D005260:Female; D005634:Fructose-Bisphosphate Aldolase / Q000096:biosynthesis / Q000235:genetics*; D007376:Interleukin-2 / Q000494:pharmacology; D007700:Kinetics; D008213:Lymphocyte Activation; D010084:Oxidation-Reduction; D011401:Promoter Regions, Genetic; D011770:Pyruvate Kinase / Q000096:biosynthesis / Q000235:genetics*; D012333:RNA, Messenger / Q000032:analysis / Q000096:biosynthesis; D051381:Rats; D017208:Rats, Wistar; D017382:Reactive Oxygen Species / Q000378:metabolism; D016329:Sp1 Transcription Factor / Q000378:metabolism*; D013601:T-Lymphocytes / Q000166:cytology / Q000276:immunology* / Q000378:metabolism*; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology; D014158:Transcription, Genetic / Q000187:drug effects
D007376:Interleukin-2; D012333:RNA, Messenger; D017382:Reactive Oxygen Species; D016329:Sp1 Transcription Factor; D003208:Concanavalin A; D011770:Pyruvate Kinase; D005634:Fructose-Bisphosphate Aldolase; D013755:Tetradecanoylphorbol Acetate