PT  - JOURNAL ARTICLE
AU  - Snijder, Joost
AU  - Schuller, Jan M.
AU  - Wiegard, Anika
AU  - Lössl, Philip
AU  - Schmelling, Nicolas
AU  - Axmann, Ilka M.
AU  - Plitzko, Jürgen M.
AU  - Förster, Friedrich
AU  - Heck, Albert J. R.
TI  - Structures of the cyanobacterial circadian oscillator frozen in a fully assembled state
AID  - 10.1126/science.aag3218
DP  - 2017 Mar 17
TA  - Science
PG  - 1181--1184
VI  - 355
IP  - 6330
4099  - http://science.sciencemag.org/content/355/6330/1181.short
4100  - http://science.sciencemag.org/content/355/6330/1181.full
SO  - Science2017 Mar 17; 355
AB  - The cyanobacterial circadian clock oscillator can be reconstituted in a test tube from just three proteins—KaiA, KaiB, and KaiC—and adenosine triphosphate (ATP). Tseng et al. studied crystal and nuclear magnetic resonance structures of complexes of the oscillator proteins and their signaling output proteins and tested the in vivo effects of structure-based mutants. Large conformational changes in KaiB and ATP hydrolysis by KaiC are coordinated with binding to output protein, which couples signaling and the day-night transitions of the clock. Snijder et al. provide complementary analysis of the oscillator proteins by mass spectrometry and cryo–electron microscopy. Their results help to explain the structural basis for the dynamic assembly of the oscillator complexes.Science, this issue p. 1174, p. 1181Cyanobacteria have a robust circadian oscillator, known as the Kai system. Reconstituted from the purified protein components KaiC, KaiB, and KaiA, it can tick autonomously in the presence of adenosine 5′-triphosphate (ATP). The KaiC hexamers enter a natural 24-hour reaction cycle of autophosphorylation and assembly with KaiB and KaiA in numerous diverse forms. We describe the preparation of stoichiometrically well-defined assemblies of KaiCB and KaiCBA, as monitored by native mass spectrometry, allowing for a structural characterization by single-particle cryo–electron microscopy and mass spectrometry. Our data reveal details of the interactions between the Kai proteins and provide a structural basis to understand periodic assembly of the protein oscillator.