Targeted transcriptional repression using a chimeric TALE-SRDX repressor protein
AuthorsMahfouz, Magdy M.
Piatek, Marek J.
Bangarusamy, Dhinoth Kumar
KAUST DepartmentBiological and Environmental Science and Engineering (BESE) Division
Bioscience Core Lab
Center for Desert Agriculture
Laboratory for Genome Engineering
Plant Stress Genomics Research Lab
Online Publication Date2011-12-14
Print Publication Date2012-02
Permanent link to this recordhttp://hdl.handle.net/10754/325247
MetadataShow full item record
AbstractTranscriptional activator-like effectors (TALEs) are proteins secreted by Xanthomonas bacteria when they infect plants. TALEs contain a modular DNA binding domain that can be easily engineered to bind any sequence of interest, and have been used to provide user-selected DNA-binding modules to generate chimeric nucleases and transcriptional activators in mammalian cells and plants. Here we report the use of TALEs to generate chimeric sequence-specific transcriptional repressors. The dHax3 TALE was used as a scaffold to provide a DNA-binding module fused to the EAR-repression domain (SRDX) to generate a chimeric repressor that targets the RD29A promoter. The dHax3. SRDX protein efficiently repressed the transcription of the RD29A
CitationMahfouz MM, Li L, Piatek M, Fang X, Mansour H, et al. (2011) Targeted transcriptional repression using a chimeric TALE-SRDX repressor protein. Plant Molecular Biology 78: 311-321. doi:10.1007/s11103-011-9866-x.
JournalPlant Molecular Biology
PubMed Central IDPMC3259320
- Dominant repression of target genes by chimeric repressors that include the EAR motif, a repression domain, in Arabidopsis.
- Authors: Hiratsu K, Matsui K, Koyama T, Ohme-Takagi M
- Issue date: 2003 Jun
- CRES-T, an effective gene silencing system utilizing chimeric repressors.
- Authors: Mitsuda N, Matsui K, Ikeda M, Nakata M, Oshima Y, Nagatoshi Y, Ohme-Takagi M
- Issue date: 2011
- A chimeric AtMYB23 repressor induces hairy roots, elongation of leaves and stems, and inhibition of the deposition of mucilage on seed coats in Arabidopsis.
- Authors: Matsui K, Hiratsu K, Koyama T, Tanaka H, Ohme-Takagi M
- Issue date: 2005 Jan
- A central regulatory system largely controls transcriptional activation and repression responses to phosphate starvation in Arabidopsis.
- Authors: Bustos R, Castrillo G, Linhares F, Puga MI, Rubio V, Pérez-Pérez J, Solano R, Leyva A, Paz-Ares J
- Issue date: 2010 Sep 9
- Synthetic TAL effectors for targeted enhancement of transgene expression in plants.
- Authors: Liu W, Rudis MR, Peng Y, Mazarei M, Millwood RJ, Yang JP, Xu W, Chesnut JD, Stewart CN Jr
- Issue date: 2014 May
Showing items related by title, author, creator and subject.
Protein implicated in nonsyndromic mental retardation regulates protein kinase A (PKA) activityAltawashi, Azza; Jung, Sung Yun; Liu, Dou; Su, Bing; Qin, Jun (Journal of Biological Chemistry, American Society for Biochemistry & Molecular Biology (ASBMB), 2012-02-28) [Article]Mutation of the coiled-coil and C2 domain-containing 1A (CC2D1A) gene, which encodes a C2 domain and DM14 domain-containing protein, has been linked to severe autosomal recessive nonsyndromic mental retardation. Using a mouse model that produces a truncated form of CC2D1A that lacks the C2 domain and three of the four DM14 domains, we show that CC2D1A is important for neuronal differentiation and brain development. CC2D1A mutant neurons are hypersensitive to stress and have a reduced capacitytoformdendritesandsynapsesinculture. Atthebiochemical level,CC2D1Atransduces signals to the cyclic adenosine 3?,5?-monophosphate (cAMP)-protein kinase A (PKA) pathway during neuronal cell differentiation. PKA activity is compromised, and the translocation of its catalytic subunit to the nucleus is also defective in CC2D1A mutant cells. Consistently, phosphorylation of the PKA target cAMP-responsive element-binding protein, at serine 133, is nearly abolished in CC2D1A mutant cells. The defects in cAMP/PKA signaling were observed in fibroblast, macrophage, and neuronal primary cells derived from the CC2D1A KO mice. CC2D1A associates with the cAMP-PKA complex following forskolin treatment and accumulates in vesicles or on the plasma membrane in wild-type cells, suggesting that CC2D1A may recruit the PKA complex to the membrane to facilitate signal transduction. Together, our data show that CC2D1A is an important regulator of the cAMP/PKA signaling pathway, which may be the underlying cause for impaired mental function in nonsyndromic mental retardation patients with CC2D1A mutation. 2012 by The American Society for Biochemistry and Molecular Biology, Inc.
Characterization of Plant Growth under Single-Wavelength Laser Light Using the Model Plant Arabidopsis ThalianaOoi, Amanda (2016-12) [Dissertation]
Advisor: Xiong, Liming
Committee members: Gehring, Christoph A; Ooi, Boon S.; Habuchi, Satoshi; Irving, HelenIndoor horticulture offers a promising solution for sustainable food production and is becoming increasingly widespread. However, it incurs high energy and cost due to the use of artificial lighting such as high-pressure sodium lamps, fluorescent light or increasingly, the light-emitting diodes (LEDs). The energy efficiency and light quality of currently available lighting is suboptimal, therefore less than ideal for sustainable and cost-effective large-scale plant production. Here, we demonstrate the use of high-powered single-wavelength lasers for indoor horticulture. Lasers are highly energy-efficient and can be remotely guided to the site of plant growth, thus reducing on-site heat accumulation. Besides, laser beams can be tailored to match the absorption profiles of different plants. We have developed a prototype laser growth chamber and demonstrate that laser-grown plants can complete a full growth cycle from seed to seed with phenotypes resembling those of plants grown under LEDs. Importantly, the plants have lower expression of proteins diagnostic for light and radiation stress. The phenotypical, biochemical and proteomic data show that the singlewavelength laser light is suitable for plant growth and therefore, potentially able to unlock the advantages of this next generation lighting technology for highly energy-efficient horticulture. Furthermore, stomatal movement partly determines the plant productivity and stress management. Abscisic acid (ABA) induces stomatal closure by promoting net K+-efflux from guard cells through outwardrectifying K+ (K+ out) channels to regulate plant water homeostasis. Here, we show that the Arabidopsis thaliana guard cell outward-rectifying K+ (ATGORK) channel is a direct target for ABA in the regulation of stomatal aperture and hence gas exchange and transpiration. Addition of (±)-ABA, but not the biologically inactive (−)-isomer, increases K+ out channel activity in Vicia faba guard cell protoplast. A similar ABA-modulated K+ channel conductance was observed when ATGORK was heterologously expressed in human embryonic kidney 293 (HEK-293) cells. Alignment of ATGORK with known PYR/PYL/RCARs ABA receptors revealed that ATGORK harbors amino acid residues that are similar to those at the latchlike region of the ABA-binding sites. In ATGORK, the double mutations K559A and Y562A at the predicted ABA-interacting site impaired ABA-dependent channel activation and reduced the affinity for ABA in vitro.