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AtMYB93 is a novel negative regulator of lateral root development in Arabidopsis.

Gibbs DJ, Voß U, Harding SA, Fannon J, Moody LA, Yamada E, Swarup K, Nibau C, Bassel GW, Choudhary A, Lavenus J, Bradshaw SJ, Stekel DJ, Bennett MJ, Coates JC - New Phytol. (2014)

Bottom Line: Furthermore, Atmyb93 mutant lateral root development is insensitive to auxin, indicating that AtMYB93 is required for normal auxin responses during lateral root development.We propose that AtMYB93 is part of a novel auxin-induced negative feedback loop stimulated in a select few endodermal cells early during lateral root development, ensuring that lateral roots only develop when absolutely required.Putative AtMYB93 homologues are detected throughout flowering plants and represent promising targets for manipulating root systems in diverse crop species.

View Article: PubMed Central - PubMed

Affiliation: School of Biosciences, University of Birmingham, Birmingham, B15 2TT, UK.

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The ARABIDILLO-1 ARMADILLO (ARM) domain interacts with three related R2R3 MYB family proteins: AtMYB92, -53 and -93. (a) Yeast two-hybrid interactions between the ARABIDILLO1 ARM-repeat domain and full-length Arabidopsis R2R3 MYB cDNAs expressed as GAL4-BD (GAL4-binding domain) and GAL4-AD (GAL4-activation domain) fusions, respectively. Growth on -LT (Leucine-Tryptophan) medium indicates successful co-transformation. Positive interactions are indicated by growth on -AHLT (Adenine-Histidine-Leucine-Tryptophan) medium and by blue colouration in the presence of X-α-gal. The ARABIDILLO-1 ARM-repeat domain interacts with AtMYB92,-53 and -93, but not with more distantly related AtMYB75/PAP1 (PAP1 = PRODUCTION OF ANTHOCYANIN PIGMENT 1) or AtMYB91/AS1 (AS1 = ASYMMETRIC LEAVES1). The interaction is specific to the C-terminus downstream of the R2R3 MYB domain. Conserved R2 and R3 MYB domains are shown in blue and magenta; the cyan box denotes the conserved C-terminal motif of AtMYB92, -53 and -93; the green box denotes a conserved C-terminal motif in AtMYB75/PAP1. (b) Co-immunoprecipitation of N-terminally MYC-tagged AtMYB92 (MYC-MYB92; closed arrowhead) with the N-terminally HA-tagged ARABIDILLO1 ARM domain (HA-ARM; open arrowhead). Proteins were synthesized in vitro, and co-incubated with anti-HA antibody. A control immunoprecipitation (IP) performed without the addition of HA-ARM was also conducted. I, input; W1/5, washes 1 and 5; E, elution; **, antibody heavy chain; *, nonspecific band in anti-MYC western blots. We performed similar experiments with AtMYB93, but because of the size of AtMYB93, it unfortunately could not be detected in the elution as it was occluded by the antibody heavy chain. (c) Alignment of the full-length amino acid sequences of AtMYB92, AtMYB53 and AtMYB93. Black and grey shading denotes identical and similar amino acid residues, respectively. Blue and magenta bars denote conserved R2 and R3 MYB domains, respectively. The cyan bar denotes the conserved C-terminal motif unique to these three proteins. *, key conserved aromatic residues within the R2R3 MYB domain.
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fig01: The ARABIDILLO-1 ARMADILLO (ARM) domain interacts with three related R2R3 MYB family proteins: AtMYB92, -53 and -93. (a) Yeast two-hybrid interactions between the ARABIDILLO1 ARM-repeat domain and full-length Arabidopsis R2R3 MYB cDNAs expressed as GAL4-BD (GAL4-binding domain) and GAL4-AD (GAL4-activation domain) fusions, respectively. Growth on -LT (Leucine-Tryptophan) medium indicates successful co-transformation. Positive interactions are indicated by growth on -AHLT (Adenine-Histidine-Leucine-Tryptophan) medium and by blue colouration in the presence of X-α-gal. The ARABIDILLO-1 ARM-repeat domain interacts with AtMYB92,-53 and -93, but not with more distantly related AtMYB75/PAP1 (PAP1 = PRODUCTION OF ANTHOCYANIN PIGMENT 1) or AtMYB91/AS1 (AS1 = ASYMMETRIC LEAVES1). The interaction is specific to the C-terminus downstream of the R2R3 MYB domain. Conserved R2 and R3 MYB domains are shown in blue and magenta; the cyan box denotes the conserved C-terminal motif of AtMYB92, -53 and -93; the green box denotes a conserved C-terminal motif in AtMYB75/PAP1. (b) Co-immunoprecipitation of N-terminally MYC-tagged AtMYB92 (MYC-MYB92; closed arrowhead) with the N-terminally HA-tagged ARABIDILLO1 ARM domain (HA-ARM; open arrowhead). Proteins were synthesized in vitro, and co-incubated with anti-HA antibody. A control immunoprecipitation (IP) performed without the addition of HA-ARM was also conducted. I, input; W1/5, washes 1 and 5; E, elution; **, antibody heavy chain; *, nonspecific band in anti-MYC western blots. We performed similar experiments with AtMYB93, but because of the size of AtMYB93, it unfortunately could not be detected in the elution as it was occluded by the antibody heavy chain. (c) Alignment of the full-length amino acid sequences of AtMYB92, AtMYB53 and AtMYB93. Black and grey shading denotes identical and similar amino acid residues, respectively. Blue and magenta bars denote conserved R2 and R3 MYB domains, respectively. The cyan bar denotes the conserved C-terminal motif unique to these three proteins. *, key conserved aromatic residues within the R2R3 MYB domain.

Mentions: The initial MYB alignment (Fig. 1c) was conducted using ClustalX (Larkin et al., 2007) using the default settings. Alignments were annotated using Boxshade 3.21 (http://www.ch.embnet.org/software/BOX_doc.html), with the fraction of sequences that must agree for shading set at 1.0. For the phylogeny, putative full-length land plant AtMYB93/92/53 homologues were identified using Blastp from fully sequenced land plant genomes via GenBank and Phytozome (Goodstein et al., 2012). Sequences were aligned using ClustalX and the alignment was refined manually in SeaView (Gouy et al., 2010). The phylogenetic tree was calculated using the maximum likelihood algorithm in SeaView on default settings, with 1000 bootstrap replicates. Similar trees were obtained using distance methods. The tree was displayed using TreeViewX (Page, 2002).


AtMYB93 is a novel negative regulator of lateral root development in Arabidopsis.

Gibbs DJ, Voß U, Harding SA, Fannon J, Moody LA, Yamada E, Swarup K, Nibau C, Bassel GW, Choudhary A, Lavenus J, Bradshaw SJ, Stekel DJ, Bennett MJ, Coates JC - New Phytol. (2014)

The ARABIDILLO-1 ARMADILLO (ARM) domain interacts with three related R2R3 MYB family proteins: AtMYB92, -53 and -93. (a) Yeast two-hybrid interactions between the ARABIDILLO1 ARM-repeat domain and full-length Arabidopsis R2R3 MYB cDNAs expressed as GAL4-BD (GAL4-binding domain) and GAL4-AD (GAL4-activation domain) fusions, respectively. Growth on -LT (Leucine-Tryptophan) medium indicates successful co-transformation. Positive interactions are indicated by growth on -AHLT (Adenine-Histidine-Leucine-Tryptophan) medium and by blue colouration in the presence of X-α-gal. The ARABIDILLO-1 ARM-repeat domain interacts with AtMYB92,-53 and -93, but not with more distantly related AtMYB75/PAP1 (PAP1 = PRODUCTION OF ANTHOCYANIN PIGMENT 1) or AtMYB91/AS1 (AS1 = ASYMMETRIC LEAVES1). The interaction is specific to the C-terminus downstream of the R2R3 MYB domain. Conserved R2 and R3 MYB domains are shown in blue and magenta; the cyan box denotes the conserved C-terminal motif of AtMYB92, -53 and -93; the green box denotes a conserved C-terminal motif in AtMYB75/PAP1. (b) Co-immunoprecipitation of N-terminally MYC-tagged AtMYB92 (MYC-MYB92; closed arrowhead) with the N-terminally HA-tagged ARABIDILLO1 ARM domain (HA-ARM; open arrowhead). Proteins were synthesized in vitro, and co-incubated with anti-HA antibody. A control immunoprecipitation (IP) performed without the addition of HA-ARM was also conducted. I, input; W1/5, washes 1 and 5; E, elution; **, antibody heavy chain; *, nonspecific band in anti-MYC western blots. We performed similar experiments with AtMYB93, but because of the size of AtMYB93, it unfortunately could not be detected in the elution as it was occluded by the antibody heavy chain. (c) Alignment of the full-length amino acid sequences of AtMYB92, AtMYB53 and AtMYB93. Black and grey shading denotes identical and similar amino acid residues, respectively. Blue and magenta bars denote conserved R2 and R3 MYB domains, respectively. The cyan bar denotes the conserved C-terminal motif unique to these three proteins. *, key conserved aromatic residues within the R2R3 MYB domain.
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Related In: Results  -  Collection

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fig01: The ARABIDILLO-1 ARMADILLO (ARM) domain interacts with three related R2R3 MYB family proteins: AtMYB92, -53 and -93. (a) Yeast two-hybrid interactions between the ARABIDILLO1 ARM-repeat domain and full-length Arabidopsis R2R3 MYB cDNAs expressed as GAL4-BD (GAL4-binding domain) and GAL4-AD (GAL4-activation domain) fusions, respectively. Growth on -LT (Leucine-Tryptophan) medium indicates successful co-transformation. Positive interactions are indicated by growth on -AHLT (Adenine-Histidine-Leucine-Tryptophan) medium and by blue colouration in the presence of X-α-gal. The ARABIDILLO-1 ARM-repeat domain interacts with AtMYB92,-53 and -93, but not with more distantly related AtMYB75/PAP1 (PAP1 = PRODUCTION OF ANTHOCYANIN PIGMENT 1) or AtMYB91/AS1 (AS1 = ASYMMETRIC LEAVES1). The interaction is specific to the C-terminus downstream of the R2R3 MYB domain. Conserved R2 and R3 MYB domains are shown in blue and magenta; the cyan box denotes the conserved C-terminal motif of AtMYB92, -53 and -93; the green box denotes a conserved C-terminal motif in AtMYB75/PAP1. (b) Co-immunoprecipitation of N-terminally MYC-tagged AtMYB92 (MYC-MYB92; closed arrowhead) with the N-terminally HA-tagged ARABIDILLO1 ARM domain (HA-ARM; open arrowhead). Proteins were synthesized in vitro, and co-incubated with anti-HA antibody. A control immunoprecipitation (IP) performed without the addition of HA-ARM was also conducted. I, input; W1/5, washes 1 and 5; E, elution; **, antibody heavy chain; *, nonspecific band in anti-MYC western blots. We performed similar experiments with AtMYB93, but because of the size of AtMYB93, it unfortunately could not be detected in the elution as it was occluded by the antibody heavy chain. (c) Alignment of the full-length amino acid sequences of AtMYB92, AtMYB53 and AtMYB93. Black and grey shading denotes identical and similar amino acid residues, respectively. Blue and magenta bars denote conserved R2 and R3 MYB domains, respectively. The cyan bar denotes the conserved C-terminal motif unique to these three proteins. *, key conserved aromatic residues within the R2R3 MYB domain.
Mentions: The initial MYB alignment (Fig. 1c) was conducted using ClustalX (Larkin et al., 2007) using the default settings. Alignments were annotated using Boxshade 3.21 (http://www.ch.embnet.org/software/BOX_doc.html), with the fraction of sequences that must agree for shading set at 1.0. For the phylogeny, putative full-length land plant AtMYB93/92/53 homologues were identified using Blastp from fully sequenced land plant genomes via GenBank and Phytozome (Goodstein et al., 2012). Sequences were aligned using ClustalX and the alignment was refined manually in SeaView (Gouy et al., 2010). The phylogenetic tree was calculated using the maximum likelihood algorithm in SeaView on default settings, with 1000 bootstrap replicates. Similar trees were obtained using distance methods. The tree was displayed using TreeViewX (Page, 2002).

Bottom Line: Furthermore, Atmyb93 mutant lateral root development is insensitive to auxin, indicating that AtMYB93 is required for normal auxin responses during lateral root development.We propose that AtMYB93 is part of a novel auxin-induced negative feedback loop stimulated in a select few endodermal cells early during lateral root development, ensuring that lateral roots only develop when absolutely required.Putative AtMYB93 homologues are detected throughout flowering plants and represent promising targets for manipulating root systems in diverse crop species.

View Article: PubMed Central - PubMed

Affiliation: School of Biosciences, University of Birmingham, Birmingham, B15 2TT, UK.

Show MeSH
Related in: MedlinePlus