title: Transactivator mutants with altered effector specificity allow selective regulation of two genes by tetracycline variants.
abstract: A set of Tet repressor (TetR) based eukaryotic transactivators that respond to 4-de(dimethylamino)-6-deoxy-6-demethyl-tetracycline (cmt3) but no longer to tetracycline (tc) is presented. The novel transactivators exhibit high activation in absence of an effector and a 200-fold reduction of reporter gene activity in the presence of cmt3. The most cmt3-sensitive mutant was coexpressed with a tc-responsive Tet transregulator harbouring an altered DNA recognition specificity. Use of cmt3 and tc yields independent control of expression of two genes in the same cell without crosstalk.
authors.affiliation: Lehrstuhl für Mikrobiologie, Friedrich-Alexander Universität Erlangen-Nürnberg, Staudtstr. 5, 91058 Erlangen, Germany.
authors.forename: Christel
authors.initials: C
authors.lastname: Krueger
authors.forename: Andreas
authors.initials: A
authors.lastname: Schmidt
authors.forename: Christina
authors.initials: C
authors.lastname: Danke
authors.forename: Wolfgang
authors.initials: W
authors.lastname: Hillen
authors.forename: Christian
authors.initials: C
authors.lastname: Berens
mesh_terms: D002455:Cell Division / Q000187:drug effects; D004305:Dose-Response Relationship, Drug; D004318:Doxycycline / Q000494:pharmacology; D005786:Gene Expression Regulation / Q000187:drug effects*; D006367:HeLa Cells; D006801:Humans; D008156:Luciferases / Q000235:genetics / Q000378:metabolism; D009154:Mutation / Q000235:genetics*; D010957:Plasmids / Q000235:genetics; D012097:Repressor Proteins / Q000235:genetics / Q000378:metabolism; D013752:Tetracycline / Q000494:pharmacology*; D013754:Tetracyclines / Q000494:pharmacology; D015534:Trans-Activators / Q000235:genetics* / Q000378:metabolism; D015533:Transcriptional Activation / Q000187:drug effects; D014162:Transfection
chemical_list: D012097:Repressor Proteins; D013754:Tetracyclines; D015534:Trans-Activators; C117155:tetracycline CMT-3; C034811:tetracycline resistance-encoding transposon repressor protein; D008156:Luciferases; D013752:Tetracycline; D004318:Doxycycline