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.
ID: qv26vTuE4
Submitter: Max Staller
Submission Date: July 10, 2018, 8:25 a.m.
Version: 1
Colors: | D | E | R | H | K | S | T | N | Q | A | V | I | L | M | F | Y | W | C | G | P |
---|
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 |
Percent Identity | Matching Chains | Protein | Accession | Entry Name |
---|---|---|---|---|
100.0 | General control protein GCN4 -Central acidic activation domain (101-144) | P03069 | GCN4_YEAST |