The p65 domain from NF-κB is an efficient human activator in the tetracycline-regulatable gene expression system

Urlinger, Stefanie; Helbl, Vera; Guthmann, J.; Pook, Elisabeth; Grimm, Silke; Hillen, Wolfgang

doi:10.1016/s0378-1119(00)00112-8

Cited by 43 publications

(33 citation statements)

References 35 publications

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“…However, this system is not efficient in mammalian cells. For this reason, and based on the knowledge acquired about how TetR binds to its target operator, several chimeric versions of TetR fused to eukaryotic regulatory domains were constructed, such as the acidic activation domain (tTA) (22,115,403) and repression domains (tTS) (33,336). Based on hybrid transregulators, transgenic mice able to produce diphtheria toxin or the regulated expression of Shiga toxin ␤ to induce apoptosis in mammalian fibroblastic cells were obtained.…”

Section: Biotechnological Applications and Future Prospectsmentioning

confidence: 99%

The TetR Family of Transcriptional Repressors

Ramos

Martínez-Bueno

Molina‐Henares

et al. 2005

Microbiol Mol Biol Rev

988

1,133

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We have developed a general profile for the proteins of the TetR family of repressors. The stretch that best defines the profile of this family is made up of 47 amino acid residues that correspond to the helix-turn-helix DNA binding motif and adjacent regions in the three-dimensional structures of TetR, QacR, CprB, and EthR, four family members for which the function and three-dimensional structure are known. We have detected a set of 2,353 nonredundant proteins belonging to this family by screening genome and protein databases with the TetR profile. Proteins of the TetR family have been found in 115 genera of gram-positive, α-, β-, and γ-proteobacteria, cyanobacteria, and archaea. The set of genes they regulate is known for 85 out of the 2,353 members of the family. These proteins are involved in the transcriptional control of multidrug efflux pumps, pathways for the biosynthesis of antibiotics, response to osmotic stress and toxic chemicals, control of catabolic pathways, differentiation processes, and pathogenicity. The regulatory network in which the family member is involved can be simple, as in TetR (i.e., TetR bound to the target operator represses tetA transcription and is released in the presence of tetracycline), or more complex, involving a series of regulatory cascades in which either the expression of the TetR family member is modulated by another regulator or the TetR family member triggers a cell response to react to environmental insults. Based on what has been learned from the cocrystals of TetR and QacR with their target operators and from their three-dimensional structures in the absence and in the presence of ligands, and based on multialignment analyses of the conserved stretch of 47 amino acids in the 2,353 TetR family members, two groups of residues have been identified. One group includes highly conserved positions involved in the proper orientation of the helix-turn-helix motif and hence seems to play a structural role. The other set of less conserved residues are involved in establishing contacts with the phosphate backbone and target bases in the operator. Information related to the TetR family of regulators has been updated in a database that can be accessed at www.bactregulators.org

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Section: Biotechnological Applications and Future Prospectsmentioning

confidence: 99%

The TetR Family of Transcriptional Repressors

Ramos

Martínez-Bueno

Molina‐Henares

et al. 2005

Microbiol Mol Biol Rev

988

1,133

View full text Add to dashboard Cite

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“…The recently reported GAL4 variant Gal4FF (Asakawa et al, 2008) utilized a dimeric form of the same minimal activation domain and is almost identical to GAL4VPmad2. Another variant used the NF-B p65 carboxyl terminus activation domain (Schmitz and Baeuerle, 1991), which has been reported to be as potent as the VP16 activation domain but less toxic (Rivera, 1998;Urlinger et al, 2000).…”

Section: Modified Gal4 Activators With Lower Toxicity Show a High Tramentioning

confidence: 99%

Adaptation of GAL4 activators for GAL4 enhancer trapping in zebrafish

Ogura

Okuda

Kondoh

et al. 2009

Developmental Dynamics

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An enhancer trap-based GAL4-UAS system in zebrafish requires strong GAL4 activators with minimal adverse effects. However, the activity of yeast GAL4 is too low in zebrafish, while a fusion protein of the GAL4 DNA-binding domain and the VP16 activation domain is toxic to embryonic development, even when expressed at low levels. To alleviate this toxicity, we developed variant GAL4 activators by fusing either multimeric forms of the VP16 minimal activation domain or the NF-B activation domain to the GAL4 DNA-binding domain. These variant GAL4 activators are sufficiently innocuous and yet highly effective transactivators in developing zebrafish. Enhancer-trap vectors containing these GAL4 activators downstream of an appropriate weak promoter were randomly inserted into the zebrafish genome using the Sleeping Beauty transposon system. By the combination of these genetic elements, we have successfully developed enhancer trap lines that activate UAS-dependent reporter genes in a tissue-specific fashion that reflects trapped enhancer activities. Developmental Dynamics 238:641-655, 2009.

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“…To overcome this problem a set of tetracycline-responsive transactivators, which are tolerated at higher intracellular concentrations, have been developed (1,5,139). Finally, remodeling of the original tTA and rtTA transactivators has been efficiently used for substantially improving several other important features of the tet system.…”

Section: The Tetracycline Systemmentioning

confidence: 99%

Of mice and models: improved animal models for biomedical research

Bockamp

Maringer

Spangenberg

et al. 2002

Physiological Genomics

147

109

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The ability to engineer the mouse genome has profoundly transformed biomedical research. During the last decade, conventional transgenic and gene knockout technologies have become invaluable experimental tools for modeling genetic disorders, assigning functions to genes, evaluating drugs and toxins, and by and large helping to answer fundamental questions in basic and applied research. In addition, the growing demand for more sophisticated murine models has also become increasingly evident. Good state-of-principle knowledge about the enormous potential of second-generation conditional mouse technology will be beneficial for any researcher interested in using these experimental tools. In this review we will focus on practice, pivotal principles, and progress in the rapidly expanding area of conditional mouse technology. The review will also present an internet compilation of available tetracycline-inducible mouse models as tools for biomedical research ( http://www.zmg.uni-mainz.de/tetmouse/ ).

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The p65 domain from NF-κB is an efficient human activator in the tetracycline-regulatable gene expression system

Cited by 43 publications

References 35 publications

The TetR Family of Transcriptional Repressors

The TetR Family of Transcriptional Repressors

Adaptation of GAL4 activators for GAL4 enhancer trapping in zebrafish

Of mice and models: improved animal models for biomedical research

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