A High-Throughput Mutational Scan of an Intrinsically Disordered Acidic Transcriptional Activation Domain.

Abstract

Transcriptional activation domains are essential for gene regulation, but their intrinsic disorder and low primary sequence conservation have made it difficult to identify the amino acid composition features that underlie their activity. Here, we describe a rational mutagenesis scheme that deconvolves the function of four activation domain sequence features-acidity, hydrophobicity, intrinsic disorder, and short linear motifs-by quantifying the activity of thousands of variants in vivo and simulating their conformational ensembles using an all-atom Monte Carlo approach. Our results with a canonical activation domain from the Saccharomyces cerevisiae transcription factor Gcn4 reconcile existing observations into a unified model of its function: the intrinsic disorder and acidic residues keep two hydrophobic motifs from driving collapse. Instead, the most-active variants keep their aromatic residues exposed to the solvent. Our results illustrate how the function of intrinsically disordered proteins can be revealed by high-throughput rational mutagenesis.

Submission Details

ID: qv26vTuE4

Submitter: Max Staller

Submission Date: July 10, 2018, 8:25 a.m.

Version: 1

Publication Details
Staller MV;Holehouse AS;Swain-Lenz D;Das RK;Pappu RV;Cohen BA,Cell Syst (2018) A High-Throughput Mutational Scan of an Intrinsically Disordered Acidic Transcriptional Activation Domain. PMID:29525204
Additional Information

Study Summary

Number of data points 21899
Proteins General control protein GCN4 -Central acidic activation domain (101-144)
Unique complexes 6340
Assays/Quantities/Protocols Experimental Assay: Activation domain variant abundance ; Experimental Assay: Activation domain induction ; Experimental Assay: Activation domain activity
Libraries Gcn4 activation domain mutants - Activity Replicate 2 ; Gcn4 activation domain mutants - Activity Replicate 1 ; Gcn4 activation domain mutants -Induction ; Gcn4 activation domain mutants - protein abundance ; Gcn4 activation domain mutants - protein abundance

Structure view and single mutant data analysis

Study data

No weblogo for data of varying length.
Colors: D E R H K S T N Q A V I L M F Y W C G P
 

Data Distribution

Studies with similar sequences (approximate matches)

Correlation with other assays (exact sequence matches)


Relevant PDB Entries

Structure ID Release Date Resolution Structure Title
2EFS 2007-02-23T00:00:00+0000 2.0 Crystal structure of the C-terminal tropomyosin fragment with N- and C-terminal extensions of the leucine zipper at 2.0 angstroms resolution
1SWI 1996-05-09T00:00:00+0000 2.6 GCN4-LEUCINE ZIPPER CORE MUTANT AS N16A COMPLEXED WITH BENZENE
1UO0 2003-09-15T00:00:00+0000 2.4 Structure Based Engineering of Internal Molecular Surfaces Of Four Helix Bundles
1UO2 2003-09-15T00:00:00+0000 1.99 Structure Based Engineering of Internal Molecular Surfaces Of Four Helix Bundles
1RB5 2003-11-01T00:00:00+0000 1.9 ANTIPARALLEL TRIMER OF GCN4-LEUCINE ZIPPER CORE MUTANT AS N16A TRIGONAL FORM
5IIV 2016-03-01T00:00:00+0000 0 GCN4p pH 1.5
1UNX 2003-09-15T00:00:00+0000 2.4 Structure Based Engineering of Internal Molecular Surfaces Of Four Helix Bundles
1UNT 2003-09-15T00:00:00+0000 2.07 Structure Based Engineering of Internal Molecular Surfaces Of Four Helix Bundles
1UO5 2003-09-15T00:00:00+0000 2.07 Structure Based Engineering of Internal Molecular Surfaces Of Four Helix Bundles
1GZL 2002-05-23T00:00:00+0000 1.8 Crystal structure of C14linkmid/IQN17: a cross-linked inhibitor of HIV-1 entry bound to the gp41 hydrophobic pocket

Relevant UniProtKB Entries

Percent Identity Matching Chains Protein Accession Entry Name
100.0 General control protein GCN4 -Central acidic activation domain (101-144) P03069 GCN4_YEAST