Regulation of triosephosphate isomerase (TPI) gene expression in TPI deficient lymphoblastoid cells.
The metabolic defect of triosephosphate isomerase (TPI) deficiency is reversible in deficient lymphoblastoid cells when cultured in the presence of human K562 erythroleukemia cells or plasma as exogenous source of functional enzyme. However, plasma contains a variety of undefined biological response modifiers whose effects on TPI gene expression are unknown. In the present study, TPI deficient lymphoblastoid cells were cultured in serum-free medium for 24 h (controls) and stimulated with fresh frozen plasma (FFP) at final concentrations of 20, 40, and 60% for 9 h. Changes in TPI mRNA expression were monitored by slot and Northern blot hybridisations using a specific human TPI cDNA probe. For equivalent loading of total RNA, TPI mRNA expression in FFP-treated lymphoblastoid cells exceeded that for controls by on average 20-fold. Additional studies with the transcription inhibitor, actinomycin D, revealed a rapid degradation of TPI mRNA in controls compared to FFP-treated cells, indicating that the stability of the TPI transcript was affected by plasma. These data suggest that functional or regulatory elements within the TPI gene promoter can be modulated by biological response modifiers. An understanding of the transcriptional control of TPI may provide useful insights into the development of gene therapy strategies for TPI deficiency.
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Department of Haematological Medicine, King's College School of Medicine and Dentistry, London SE5 9RS, UK.
Humphries
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Layton
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D002460:Cell Line; D016895:Culture Media, Serum-Free / Q000494:pharmacology; D003609:Dactinomycin / Q000494:pharmacology; D015971:Gene Expression Regulation, Enzymologic; D005796:Genes / Q000235:genetics*; D006801:Humans; D008214:Lymphocytes / Q000166:cytology / Q000187:drug effects / Q000201:enzymology; D019384:Nucleic Acid Synthesis Inhibitors / Q000494:pharmacology; D010949:Plasma; D012333:RNA, Messenger / Q000378:metabolism; D014158:Transcription, Genetic / Q000187:drug effects; D014305:Triose-Phosphate Isomerase / Q000235:genetics*
D016895:Culture Media, Serum-Free; D019384:Nucleic Acid Synthesis Inhibitors; D012333:RNA, Messenger; D003609:Dactinomycin; D014305:Triose-Phosphate Isomerase