Comprehensive identification of <i>Arabidopsis thaliana</i> MYB transcription factors interacting with R/B‐like BHLH proteins

Zimmermann, Ilona; Heim, Marc A.; Weißhaar, Bernd; Uhrig, Joachim F.

doi:10.1111/j.1365-313x.2004.02183.x

Cited by 756 publications

(804 citation statements)

References 58 publications

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“…In accordance with published data 12 interaction between MYB and BHLH transcription factor family members (CPC and GL3, respectively; see Supplementary Methods and Supplementary Fig. 2 online) could be detected by a red fluorescent signal in the nucleus (Fig.…”

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confidence: 91%

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An improved mRFP1 adds red to bimolecular fluorescence complementation

et al. 2006

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Protein-protein interactions are fundamental to virtually every aspect of cellular functions. Blue, green and yellow bimolecular fluorescence complementation (BiFC) systems based on GFP and its variants allow the investigation of protein-protein interactions in vivo. We have developed the first red BiFC system based on an improved monomeric red fluorescent protein (mRFP1-Q66T), expanding the range of possible applications for BiFC.With interactome data available for several model organisms, a challenging next step in post-genomic research is to analyze protein complex formation in vivo. The recently developed BiFC assay is a comparably fast and simple noninvasive technology to study protein interactions inside living cells. BiFC is based on the reconstitution of the yellow fluorescent protein (YFP) from two nonfluorescent fragments when they are brought into close proximity by a physical interaction between proteins fused to each fragment 1 . This approach has proven to be robust and versatile, and multicolor versions using spectral variants of GFP further increase the potential of this technology 2 . The color spectrum available for BiFC, however, has been limited to blue, green and yellow.The red fluorescent protein from Discosoma sp. (DsRED) and its variants are established intracellular reporter proteins, but the characteristics of most DsRED variants, particularly their obligate tetramerization, impede their application in BiFC 3,4 . An extensively mutated monomeric DsRED variant (mRFP1) has been generated 5 , but substantially altered spectra, poor brightness and low photostability limit its usefulness as a reporter protein.We have identified improved mRFP1 mutants, which allowed us to establish a red fluorescent reporter system for detection of protein interactions in vivo. We used site-directed mutagenesis to generate mRFP1 variants with a randomized first position of the fluorophore. Screening of 5,000 colonies resulted in the identification of 50 clones exhibiting strong red fluorescence representing three different mutations (Q66C, Q66S and Q66T) with frequencies of 5%, 43% and 52%, respectively.

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confidence: 91%

“…2e,f). As a negative control, we used CPC containing a single amino acid substitution (R63D) in the MYB domain, shown previously to specifically disrupt the interaction 12 . Transformed cells exhibited no detectable red fluorescence (Fig.…”

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confidence: 99%

An improved mRFP1 adds red to bimolecular fluorescence complementation

et al. 2006

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“…Rather, the transcription factor may induce the expression of a group of specific target genes, either through saltinduced structural modification or by working cooperatively with other salt-activated transcription factors. In many cases, MYB transcription factors interact with basic helix-loop-helix (bHLH) transcription factors to exert their specific roles (Grotewold et al, 2000;Zimmermann et al, 2004;Quattrocchio et al, 2006). Further studies to identify the target genes, binding sites on promoters, and interacting proteins would clearly define the biological roles of the AtMYB44 transcription factor.…”

Section: Discussionmentioning

confidence: 99%

Overexpression of AtMYB44 Enhances Stomatal Closure to Confer Abiotic Stress Tolerance in Transgenic Arabidopsis

et al. 2007

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AtMYB44 belongs to the R2R3 MYB subgroup 22 transcription factor family in Arabidopsis (Arabidopsis thaliana). Treatment with abscisic acid (ABA) induced AtMYB44 transcript accumulation within 30 min. The gene was also activated under various abiotic stresses, such as dehydration, low temperature, and salinity. In transgenic Arabidopsis carrying an AtMYB44 promoterdriven b-glucuronidase (GUS) construct, strong GUS activity was observed in the vasculature and leaf epidermal guard cells. Transgenic Arabidopsis overexpressing AtMYB44 is more sensitive to ABA and has a more rapid ABA-induced stomatal closure response than wild-type and atmyb44 knockout plants. Transgenic plants exhibited a reduced rate of water loss, as measured by the fresh-weight loss of detached shoots, and remarkably enhanced tolerance to drought and salt stress compared to wild-type plants. Microarray analysis and northern blots revealed that salt-induced activation of the genes that encode a group of serine/threonine protein phosphatases 2C (PP2Cs), such as ABI1, ABI2, AtPP2CA, HAB1, and HAB2, was diminished in transgenic plants overexpressing AtMYB44. By contrast, the atmyb44 knockout mutant line exhibited enhanced salt-induced expression of PP2C-encoding genes and reduced drought/salt stress tolerance compared to wild-type plants. Therefore, enhanced abiotic stress tolerance of transgenic Arabidopsis overexpressing AtMYB44 was conferred by reduced expression of genes encoding PP2Cs, which have been described as negative regulators of ABA signaling.

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“…Our results, however, suggest that JA signaling activates the function of GL1 protein in an indirect manner. Several R2R3-Myb proteins, including GL1, share a conserved motif within their Myb domains that physically interacts with R/B-like bHLH proteins (Grotewold et al, 2000;Zimmermann et al, 2004). The gl1-S92F mutant, in which the Myb domain of GL1 is disrupted (Ser92Phe substitution), did not induce any trichomes in response to wounding or MeJA treatment ( Fig.…”

Section: Discrete Domains Of Gl1 Contribute To Trichome Induction Difmentioning

confidence: 99%

Jasmonic acid control of GLABRA3 links inducible defense and trichome patterning inArabidopsis

et al. 2009

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Once attacked by herbivores, plants regenerate new leaves with increased trichome density as an inducible defense. Trichomes are specified from neighboring epidermal cells through local cell-cell interactions in the leaf primordia. However, the molecular mechanism of how herbivore-induced damage at older leaves remodels the pattern of trichome fate specification at newly forming leaves is largely unknown. In this study, we show that mutations in either the biosynthetic or signaling pathway of jasmonates (JAs), long-distance wound signals, abolish the wound-induced formation of trichomes. To identify the factors linking JA signaling to trichome fate specification, we isolated a novel class of mutants, unarmed (urm), which lack trichome induction but show otherwise normal responses to JAs. URM9 encodes an Importin β family protein, and URM23 is identical to TRANSPARENT TESTA GLABRA1 (TTG1), the product of which interacts with the bHLH transcription factor GLABRA3 (GL3). Loss of either URM9 or URM23 disrupts the subnuclear localization of GL3, thus implicating GL3 in trichome induction. The expression of GL3 was enhanced by JA treatment prior to trichome initiation. Genetic analysis of multiple trichome mutants shows that GL3, in concert with the R2R3-Myb transcription factor GLABRA1 (GL1), promotes trichome fate in response to JA in a dosage-dependent manner. These results indicate that GL3 is a key transcription factor of wound-induced trichome formation acting downstream of JA signaling in Arabidopsis.

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Comprehensive identification of Arabidopsis thaliana MYB transcription factors interacting with R/B‐like BHLH proteins

Cited by 756 publications

References 58 publications

An improved mRFP1 adds red to bimolecular fluorescence complementation

An improved mRFP1 adds red to bimolecular fluorescence complementation

Overexpression of AtMYB44 Enhances Stomatal Closure to Confer Abiotic Stress Tolerance in Transgenic Arabidopsis

Jasmonic acid control of GLABRA3 links inducible defense and trichome patterning inArabidopsis

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