Mediator Complex Subunits MED2, MED5, MED16, and MED23 Genetically Interact in the Regulation of Phenylpropanoid Biosynthesis

Dolan, Whitney L.; Dilkes, Brian P.; Stout, Jake M.; Bonawitz, Nicholas D.; Chapple, Clint

doi:10.1105/tpc.17.00282

Title: Mediator Complex Subunits MED2, MED5, MED16, and MED23 Genetically Interact in the Regulation of Phenylpropanoid Biosynthesis

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Abstract

The phenylpropanoid pathway is a major global carbon sink and is important for plant fitness and the engineering of bioenergy feedstocks. In Arabidopsis thaliana, disruption of two subunits of the transcriptional regulatory Mediator complex, MED5a and MED5b, results in an increase in phenylpropanoid accumulation. By contrast, the semidominant MED5b mutation reduced epidermal fluorescence4-3 (ref4-3) results in dwarfism and constitutively repressed phenylpropanoid accumulation. Here, we report the results of a forward genetic screen for suppressors of ref4-3. We identified 13 independent lines that restore growth and/or phenylpropanoid accumulation in the ref4-3 background. Two of the suppressors restore growth without restoring soluble phenylpropanoid accumulation, indicating that the growth and metabolic phenotypes of the ref4-3 mutant can be genetically disentangled. Whole-genome sequencing revealed that all but one of the suppressors carry mutations in MED5b or other Mediator subunits. RNA-seq analysis showed that the ref4-3 mutation causes widespread changes in gene expression, including the upregulation of negative regulators of the phenylpropanoid pathway, and that the suppressors reverse many of these changes. Together, our data highlight the interdependence of individual Mediator subunits and provide greater insight into the transcriptional regulation of phenylpropanoid biosynthesis by the Mediator complex.

Authors:

^[1];

^[1]; Bonawitz, Nicholas D. ^[1];

^[1]

Department of Biochemistry, Purdue University, West Lafayette, Indiana 47907, Purdue Center for Plant Biology, West Lafayette, Indiana 47907

Publication Date:: Mon Dec 04 00:00:00 EST 2017

Research Org.:: Purdue Univ., West Lafayette, IN (United States)

Sponsoring Org.:: USDOE Office of Science (SC), Basic Energy Sciences (BES). Chemical Sciences, Geosciences, and Biosciences Division

OSTI Identifier:: 1411117

Alternate Identifier(s):: OSTI ID: 1597887

Grant/Contract Number:: FG02-07ER15905

Resource Type:: Published Article

Journal Name:: The Plant Cell

Additional Journal Information:: Journal Name: The Plant Cell Journal Volume: 29 Journal Issue: 12; Journal ID: ISSN 1040-4651

Publisher:: Oxford University Press

Country of Publication:: United States

Language:: English

Subject:: 09 BIOMASS FUELS

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                    Dolan, Whitney L., Dilkes, Brian P., Stout, Jake M., Bonawitz, Nicholas D., and Chapple, Clint. Mediator Complex Subunits MED2, MED5, MED16, and MED23 Genetically Interact in the Regulation of Phenylpropanoid Biosynthesis.  United States: N. p., 2017. 
Web.  doi:10.1105/tpc.17.00282.

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                    Dolan, Whitney L., Dilkes, Brian P., Stout, Jake M., Bonawitz, Nicholas D., & Chapple, Clint. Mediator Complex Subunits MED2, MED5, MED16, and MED23 Genetically Interact in the Regulation of Phenylpropanoid Biosynthesis.  United States.  https://doi.org/10.1105/tpc.17.00282

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                    Dolan, Whitney L., Dilkes, Brian P., Stout, Jake M., Bonawitz, Nicholas D., and Chapple, Clint. Mon .  
"Mediator Complex Subunits MED2, MED5, MED16, and MED23 Genetically Interact in the Regulation of Phenylpropanoid Biosynthesis".  United States.  https://doi.org/10.1105/tpc.17.00282.

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@article{osti_1411117,

  title        = {Mediator Complex Subunits MED2, MED5, MED16, and MED23 Genetically Interact in the Regulation of Phenylpropanoid Biosynthesis},

  author       = {Dolan, Whitney L. and Dilkes, Brian P. and Stout, Jake M. and Bonawitz, Nicholas D. and Chapple, Clint},

  abstractNote = {The phenylpropanoid pathway is a major global carbon sink and is important for plant fitness and the engineering of bioenergy feedstocks. In Arabidopsis thaliana, disruption of two subunits of the transcriptional regulatory Mediator complex, MED5a and MED5b, results in an increase in phenylpropanoid accumulation. By contrast, the semidominant MED5b mutation reduced epidermal fluorescence4-3 (ref4-3) results in dwarfism and constitutively repressed phenylpropanoid accumulation. Here, we report the results of a forward genetic screen for suppressors of ref4-3. We identified 13 independent lines that restore growth and/or phenylpropanoid accumulation in the ref4-3 background. Two of the suppressors restore growth without restoring soluble phenylpropanoid accumulation, indicating that the growth and metabolic phenotypes of the ref4-3 mutant can be genetically disentangled. Whole-genome sequencing revealed that all but one of the suppressors carry mutations in MED5b or other Mediator subunits. RNA-seq analysis showed that the ref4-3 mutation causes widespread changes in gene expression, including the upregulation of negative regulators of the phenylpropanoid pathway, and that the suppressors reverse many of these changes. Together, our data highlight the interdependence of individual Mediator subunits and provide greater insight into the transcriptional regulation of phenylpropanoid biosynthesis by the Mediator complex.},

  doi          = {10.1105/tpc.17.00282},

  journal      = {The Plant Cell},

  number       = 12,

  volume       = 29,

  place        = {United States},

  year         = {Mon Dec 04 00:00:00 EST 2017},

  month        = {Mon Dec 04 00:00:00 EST 2017}

}

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