Abstract
The plant corepressor TOPLESS (TPL) is recruited to a large number of loci that are selectively induced in response to developmental or environmental cues, yet the mechanisms by which it inhibits expression in the absence of these stimuli are poorly understood. Previously, we had used the N-terminus of Arabidopsis thaliana TPL to enable repression of a synthetic auxin response circuit in Saccharomyces cerevisiae (yeast). Here, we leveraged the yeast system to interrogate the relationship between TPL structure and function, specifically scanning for repression domains. We identified a potent repression domain in Helix 8 located within the CRA domain, which directly interacted with the Mediator middle module subunits Med21 and Med10. Interactions between TPL and Mediator were required to fully repress transcription in both yeast and plants. In contrast, we found that multimer formation, a conserved feature of many corepressors, had minimal influence on the repression strength of TPL.
Keywords:
A. thaliana; S. cerevisiae; TOPLESS; chromosomes; corepressors; gene expression; mediator; plant biology; transcriptional repression.
© 2021, Leydon et al.
Publication types
-
Research Support, N.I.H., Extramural
-
Research Support, Non-U.S. Gov't
MeSH terms
-
Arabidopsis / genetics
-
Arabidopsis / metabolism*
-
Arabidopsis Proteins / chemistry
-
Arabidopsis Proteins / genetics
-
Arabidopsis Proteins / metabolism*
-
Co-Repressor Proteins / chemistry
-
Co-Repressor Proteins / genetics
-
Co-Repressor Proteins / metabolism*
-
Gene Expression Regulation, Fungal
-
Gene Expression Regulation, Plant
-
Mediator Complex / chemistry
-
Mediator Complex / genetics
-
Mediator Complex / metabolism*
-
Models, Molecular
-
Nuclear Proteins / genetics
-
Nuclear Proteins / metabolism
-
Plants, Genetically Modified / genetics
-
Plants, Genetically Modified / metabolism*
-
Protein Binding
-
Protein Conformation, alpha-Helical
-
Protein Interaction Domains and Motifs
-
Repressor Proteins / genetics
-
Repressor Proteins / metabolism
-
Saccharomyces cerevisiae / genetics
-
Saccharomyces cerevisiae / metabolism
-
Saccharomyces cerevisiae Proteins / genetics
-
Saccharomyces cerevisiae Proteins / metabolism
-
Structure-Activity Relationship
Substances
-
Arabidopsis Proteins
-
Co-Repressor Proteins
-
Mediator Complex
-
NUT2 protein, S cerevisiae
-
Nuclear Proteins
-
Repressor Proteins
-
Saccharomyces cerevisiae Proteins
-
TUP1 protein, S cerevisiae
-
topless protein, Arabidopsis