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

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
1CE9 1999-03-18T00:00:00+0000 1.8 HELIX CAPPING IN THE GCN4 LEUCINE ZIPPER
1DGC 1993-07-15T00:00:00+0000 3.0 THE X-RAY STRUCTURE OF THE GCN4-BZIP BOUND TO ATF/CREB SITE DNA SHOWS THE COMPLEX DEPENDS ON DNA FLEXIBILITY
1FAV 2000-07-13T00:00:00+0000 3.0 THE STRUCTURE OF AN HIV-1 SPECIFIC CELL ENTRY INHIBITOR IN COMPLEX WITH THE HIV-1 GP41 TRIMERIC CORE
1FMH 2000-08-17T00:00:00+0000 0 NMR SOLUTION STRUCTURE OF A DESIGNED HETERODIMERIC LEUCINE ZIPPER
1FMH 2000-08-17T00:00:00+0000 0 NMR SOLUTION STRUCTURE OF A DESIGNED HETERODIMERIC LEUCINE ZIPPER
1GCL 1993-10-20T00:00:00+0000 2.1 GCN4 LEUCINE ZIPPER CORE MUTANT P-LI
1GCM 1995-04-25T00:00:00+0000 1.8 GCN4 LEUCINE ZIPPER CORE MUTANT P-LI
1GK6 2001-08-08T00:00:00+0000 1.9 Human vimentin coil 2B fragment linked to GCN4 leucine zipper (Z2B)
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
1IHQ 2001-04-19T00:00:00+0000 0 GLYTM1BZIP: A CHIMERIC PEPTIDE MODEL OF THE N-TERMINUS OF A RAT SHORT ALPHA TROPOMYOSIN WITH THE N-TERMINUS ENCODED BY EXON 1B

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