Casein Kinase 1 Underlies Temperature Compensation of Circadian Rhythms in Human Red Blood Cells

Abstract

Temperature compensation and period determination by casein kinase 1 (CK1) are conserved features of eukaryotic circadian rhythms, whereas the clock gene transcription factors that facilitate daily gene expression rhythms differ between phylogenetic kingdoms. Human red blood cells (RBCs) exhibit temperature-compensated circadian rhythms, which, because RBCs lack nuclei, must occur in the absence of a circadian transcription-translation feedback loop. We tested whether period determination and temperature compensation are dependent on CKs in RBCs. As with nucleated cell types, broad-spectrum kinase inhibition with staurosporine lengthened the period of the RBC clock at 37°C, with more specific inhibition of CK1 and CK2 also eliciting robust changes in circadian period. Strikingly, inhibition of CK1 abolished temperature compensation and increased the Q 10 for the period of oscillation in RBCs, similar to observations in nucleated cells. This indicates that CK1 activity is essential for circadian rhythms irrespective of the presence or absence of clock gene expression cycles.

CRYPTOCHROMES confer robustness, not rhythmicity, to circadian timekeeping

“…In this context, CK1δ/ε frequently operates in tandem with glycogen synthase kinase (GSK) 3α/β, as occurs in the regulation of β‐catenin stability (O’Neill et al , 2013; Robertson et al , 2018). Interestingly, both CK1δ/ε and GSK3α/β have a conserved role in determining the speed at which the eukaryotic cellular circadian clock runs (Hastings et al , 2008; Causton et al , 2015), both in the presence and absence of transcription (Hirota et al , 2008; Meng et al , 2008; O’Neill et al , 2011; Beale et al , 2019). This is despite the fact that the clock proteins phosphorylated by these kinases are highly dissimilar between animals, plants and fungi (Causton et al , 2015; Wong & O’Neill, 2018).…”

Ten–Second Electrophysiology: Evaluation of the 3DEP Platform for high-speed, high-accuracy cell analysis

“…It has been suggested that this surface component could be significantly larger than the conventional whole-cell parameter, particularly for smaller cells, making it difficult to extract the membrane conductance parameter. However, this can still be used as an effective discriminator of cellular properties and has been used to detect changes in RBC membrane properties due to malarial infection 49 , as well as identifying circadian rhythms in RBC electrophysiology 25,26 . For Jurkat cells a value of 11.3 nS (equivalent to a specific membrane resistance of 0.45 kΩ.cm 2 ) was obtained which is larger than common values of whole-cell conductance reported for other cell types of similar origin by patch-clamp, though by less than an order of magnitude.…”

“…DEP has been used to measure electrical properties of excitable cells and benchmarks well against other, validated methods 18 . DEP-spectra have also successfully defined biomarkers for stem cell differentiation 5,19 , cancer 20,21 , drug interactions 22–24 and circadian biology 25,26 . However, although DEP is most effective for measuring cell electrical properties, its adoption as a method in general cell biology has been limited by electrode technology and the challenges associated with robust data analysis.…”

Review: Dielectrophoresis in cell characterization

“…Perhaps most intriguing, this difference was found to be a function of, Raman spectroscopy confirmed, membrane morphology and composition differences rather than cell size [128]. Our group also demonstrated DEP's corroboration with FACS and inductively coupled plasma mass spectrometry (ICPMS) in the investigations of circadian rhythms in RBCs [126, 129]. Adams et al demonstrated that one particular DEP‐based cell sorter, Dielectrophoresis assisted continuous sorter (DACS), had even better NP enrichment than FACS of neuron NPs [86–87].…”