RRC ID |
53813
|
著者 |
Shinohara Y, Koyama YM, Ukai-Tadenuma M, Hirokawa T, Kikuchi M, Yamada RG, Ukai H, Fujishima H, Umehara T, Tainaka K, Ueda HR.
|
タイトル |
Temperature-Sensitive Substrate and Product Binding Underlie Temperature-Compensated Phosphorylation in the Clock.
|
ジャーナル |
Mol Cell
|
Abstract |
Temperature compensation is a striking feature of the circadian clock. Here we investigate biochemical mechanisms underlying temperature-compensated, CKIδ-dependent multi-site phosphorylation in mammals. We identify two mechanisms for temperature-insensitive phosphorylation at higher temperature: lower substrate affinity to CKIδ-ATP complex and higher product affinity to CKIδ-ADP complex. Inhibitor screening of ADP-dependent phosphatase activity of CKIδ identified aurintricarboxylic acid (ATA) as a temperature-sensitive kinase activator. Docking simulation of ATA and mutagenesis experiment revealed K224D/K224E mutations in CKIδ that impaired product binding and temperature-compensated primed phosphorylation. Importantly, K224D mutation shortens behavioral circadian rhythms and changes the temperature dependency of SCN's circadian period. Interestingly, temperature-compensated phosphorylation was evolutionary conserved in yeast. Molecular dynamics simulation and X-ray crystallography demonstrate that an evolutionally conserved CKI-specific domain around K224 can provide a structural basis for temperature-sensitive substrate and product binding. Surprisingly, this domain can confer temperature compensation on a temperature-sensitive TTBK1. These findings suggest the temperature-sensitive substrate- and product-binding mechanisms underlie temperature compensation.
|
巻・号 |
67(5)
|
ページ |
783-798.e20
|
公開日 |
2017-9-7
|
DOI |
10.1016/j.molcel.2017.08.009
|
PII |
S1097-2765(17)30606-8
|
PMID |
28886336
|
MeSH |
Adenosine Triphosphate / metabolism*
Animals
Binding Sites
Casein Kinase Idelta / chemistry
Casein Kinase Idelta / genetics
Casein Kinase Idelta / metabolism*
Catalytic Domain
Circadian Clocks*
Circadian Rhythm*
Crystallography, X-Ray
Genotype
HEK293 Cells
Humans
Hydrolysis
Kinetics
Locomotion
Mice, Transgenic
Models, Biological
Molecular Docking Simulation
Molecular Dynamics Simulation
Mutation
Phenotype
Phosphorylation
Protein Binding
Protein Domains
Saccharomyces cerevisiae / enzymology
Saccharomyces cerevisiae / genetics
Serine
Structure-Activity Relationship
Substrate Specificity
Suprachiasmatic Nucleus / enzymology*
Temperature*
Tissue Culture Techniques
Transfection
|
IF |
15.584
|
引用数 |
26
|
リソース情報 |
遺伝子材料 |
pHisGST CKI delta (RDB15964)
pHisGST CKI homolog 1 (RDB15965) |