Sequence Determinants of the Conformational Properties of an Intrinsically Disordered Protein Prior to and upon Multisite Phosphorylation.
Many cell signaling events are coordinated by intrinsically disordered protein regions (IDRs) that undergo multisite Serine/Threonine phosphorylation. The conformational properties of these IDRs prior to and following multisite phosphorylation are directly relevant to understanding their functions. Here, we present results from biophysical studies and molecular simulations that quantify the conformational properties of an 81-residue IDR from the S. cerevisiae transcription factor Ash1. We show that the unphosphorylated Ash1 IDR adopts coil-like conformations that are expanded and well-solvated. This result contradicts inferences regarding global compaction that are derived from heuristics based on amino acid compositions for IDRs with low proline contents. Upon phosphorylation at ten distinct sites, the global conformational properties of pAsh1 are indistinguishable from those of unphosphorylated Ash1. This insensitivity derives from compensatory changes to the pattern of local and long-range intrachain contacts. We show that the conformational properties of Ash1 and pAsh1 can be explained in terms of the linear sequence patterning of proline and charged residues vis-à-vis all other residues. The sequence features of the Ash1 IDR are shared by many other IDRs that undergo multisite phosphorylation. Accordingly, we propose that our findings might be generalizable to other IDRs involved in cell signaling.
Department of Structural Biology, St. Jude Children's Research Hospital , 263 Danny Thomas Place, Memphis, Tennessee 38105, United States.
Erik W
EW
Martin
Department of Biomedical Engineering and Center for Biological Systems Engineering, Washington University in St. Louis , One Brookings Drive, Campus Box 1097, St. Louis, Missouri 63130, United States.
Alex S
AS
Holehouse
Department of Structural Biology, St. Jude Children's Research Hospital , 263 Danny Thomas Place, Memphis, Tennessee 38105, United States.
Christy R
CR
Grace
Department of Structural Biology, St. Jude Children's Research Hospital , 263 Danny Thomas Place, Memphis, Tennessee 38105, United States.
Alex
A
Hughes
Department of Biomedical Engineering and Center for Biological Systems Engineering, Washington University in St. Louis , One Brookings Drive, Campus Box 1097, St. Louis, Missouri 63130, United States.
Rohit V
RV
Pappu
Department of Structural Biology, St. Jude Children's Research Hospital , 263 Danny Thomas Place, Memphis, Tennessee 38105, United States.
Tanja
T
Mittag
D000595:Amino Acid Sequence; D064267:Intrinsically Disordered Proteins / Q000737:chemistry* / Q000378:metabolism*; D010766:Phosphorylation; D011487:Protein Conformation; D012441:Saccharomyces cerevisiae / Q000737:chemistry* / Q000378:metabolism
D064267:Intrinsically Disordered Proteins