Control of transcriptional activity by design of charge patterning in the intrinsically disordered RAM region of the Notch receptor
| Autor: | Doug Barrick, Kathryn P. Sherry, Rohit V. Pappu, Rahul K. Das |
|---|---|
| Rok vydání: | 2017 |
| Předmět: |
0301 basic medicine
Ankyrins Transcriptional Activation Transcription Genetic Notch signaling pathway Plasma protein binding Biology Intrinsically disordered proteins DNA-binding protein 03 medical and health sciences Ankyrin Humans Amino Acid Sequence Peptide sequence Transcription factor chemistry.chemical_classification Multidisciplinary 030102 biochemistry & molecular biology Receptors Notch Protein Structure Tertiary DNA-Binding Proteins Intrinsically Disordered Proteins Crystallography 030104 developmental biology chemistry PNAS Plus Biophysics Ankyrin repeat Protein Binding Signal Transduction |
| Zdroj: | Proceedings of the National Academy of Sciences of the United States of America. 114(44) |
| ISSN: | 1091-6490 |
| Popis: | Intrinsically disordered regions (IDRs) play important roles in proteins that regulate gene expression. A prominent example is the intracellular domain of the Notch receptor (NICD), which regulates the transcription of Notch-responsive genes. The NICD sequence includes an intrinsically disordered RAM region and a conserved ankyrin (ANK) domain. The 111-residue RAM region mediates bivalent interactions of NICD with the transcription factor CSL. Although the sequence of RAM is poorly conserved, the linear patterning of oppositely charged residues shows minimal variation. The conformational properties of polyampholytic IDRs are governed as much by linear charge patterning as by overall charge content. Here, we used sequence design to assess how changing the charge patterning within RAM affects its conformational properties, the affinity of NICD to CSL, and Notch transcriptional activity. Increased segregation of oppositely charged residues leads to linear decreases in the global dimensions of RAM and decreases the affinity of a construct including a C-terminal ANK domain (RAMANK) for CSL. Increasing charge segregation from WT RAM sharply decreases transcriptional activation for all permutants. Activation also decreases for some, but not all, permutants with low charge segregation, although there is considerable variation. Our results suggest that the RAM linker is more than a passive tether, contributing local and/or long-range sequence features that modulate interactions within NICD and with downstream components of the Notch pathway. We propose that sequence features within IDRs have evolved to ensure an optimal balance of sequence-encoded conformational properties, interaction strengths, and cellular activities. |
| Databáze: | OpenAIRE |
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