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Single-chain Tet transregulators.

TitleSingle-chain Tet transregulators.
Publication TypeJournal Article
Year of Publication2003
AuthorsKrueger C, Berens C, Schmidt A, Schnappinger D, Hillen W
JournalNucleic Acids Res
Volume31
Issue12
Pagination3050-6
Date Published2003 Jun 15
ISSN1362-4962
KeywordsCell Line, Dimerization, Escherichia coli, Gene Silencing, HeLa Cells, Humans, Protein Structure, Tertiary, Recombinant Fusion Proteins, Repressor Proteins, Trans-Activators
Abstract

We demonstrate here that the Tet repressor (TetR), a dimeric allosterical regulatory protein, can be converted to a fully functional monomer when connected by a 29 amino acid linker. TetR-based transregulators are widely used to regulate gene expression in eukaryotes. They can be fused to form single-chain (sc) Tet transregulators with two TetR moieties and one eukaryotic regulatory domain. Sc variants of transactivator and transsilencer exhibit the same regulatory properties as their respective dimeric counterparts in human cell lines. In particular, the reverse 'tet-on' phenotype of rtTA variants is also present in the sc variants. Coexpression of a reverse transactivator and sc transsilencer leads to reduced background expression and shows full activation upon induction. The data demonstrate that sc Tet transregulators exhibit the phenotype of their respective dimers and lack functional interference when coexpressed in the same cell.

DOI10.1093/nar/gkg421
Alternate JournalNucleic Acids Res
PubMed ID12799431
PubMed Central IDPMC162254

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