Lack of gradual regulation of tetracycline-controlled gene expression by the tetracyclin-repressor/VP16 transactivator (tTA) in cultured cells.
Von Willebrand factor (vWF) is an essential multimeric protein for adhesion of platelets to an injured vessel wall. Endothelial cells secrete vWF by either a constitutive or a regulated pathway. It is unknown whether the secretory partitioning of vWF is dependent on the level of vWF synthesis. We employed the widely applied tetracycline-controlled transactivator system (tTA) to study the regulation of vWF mRNA synthesis in stably transfected Madin Darby kidney (MDCK-II) cells in a quantitative manner. Immunofluorescence staining with anti-vWF antibodies revealed that increasing the concentration of tetracycline resulted in a decreased number of MDCK-II cells that synthesize vWF. Apparently, tTA-regulated gene expression in an individual cell functions as an 'on/off' system rather than regulating the level of gene expression in a dose-response manner, as reported previously.
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Department of Biochemistry, University of Amsterdam, The Netherlands.
de Waard
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V
van Mourik
J A
JA
Pannekoek
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H
D000818:Animals; D000900:Anti-Bacterial Agents / Q000494:pharmacology; D002451:Cell Compartmentation; D002478:Cells, Cultured; D004285:Dogs; D004305:Dose-Response Relationship, Drug; D005786:Gene Expression Regulation / Q000187:drug effects; D017405:Herpes Simplex Virus Protein Vmw65 / Q000235:genetics; D007668:Kidney / Q000166:cytology; D011994:Recombinant Proteins; D012097:Repressor Proteins / Q000235:genetics; D013752:Tetracycline / Q000494:pharmacology; D014162:Transfection; D014841:von Willebrand Factor / Q000096:biosynthesis* / Q000235:genetics
D000900:Anti-Bacterial Agents; D017405:Herpes Simplex Virus Protein Vmw65; D011994:Recombinant Proteins; D012097:Repressor Proteins; C034811:tetracycline resistance-encoding transposon repressor protein; D014841:von Willebrand Factor; D013752:Tetracycline