An integrative omics approach reveals posttranscriptional mechanisms underlying circadian temperature compensation

Schmal, C.; Maier, B.; Ashwal-Fluss, R.; Bartok, O.; Finger, A.-M.; Bange, T.; Koutsouli, S.; Robles, M. S.; Kadener, S.; Herzel, H.; Kramer, A.

doi:10.1101/2021.10.06.463236

bioRxiv · 10.1101/2021.10.06.463236

An integrative omics approach reveals posttranscriptional mechanisms underlying circadian temperature compensation

Schmal, C.·Maier, B.·Ashwal-Fluss, R.·Bartok, O.·Finger, A.-M.·Bange, T.·Koutsouli, S.·Robles, M. S.·Kadener, S.·Herzel, H.·Kramer, A.

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Abstract

A defining property of circadian clocks is temperature compensation, characterized by the resilience of circadian free-running periods against changes in environmental temperature. As an underlying mechanism, the balance or critical reaction hypothesis have been proposed. While the former supposes a temperature-dependent balancing of reactions with opposite effects on circadian period, the latter assumes an insensitivity of certain critical period determining regulations upon temperature changes. Posttranscriptional regulations such as temperature-sensitive alternative splicing or phosphorylation have been described as underlying reactions. Here, we show that knockdown of cleavage and polyadenylation specificity factor subunit 6 (CPSF6), a key regulator of 3-end cleavage and polyadenylation, abolishes circadian temperature compensation in U-2 OS cells. We apply a combination of 3-End-RNA-seq and mass spectrometry-based proteomics to globally quantify changes in 3 UTR length as well as gene and protein expression between wild type and CPSF6 knock-down cells and their dependency on temperature. Analyzing differential responses upon temperature changes in wild type and CPSF6 knockdown cells reveals candidate genes underlying circadian temperature compensation. We identify that eukaryotic translation initiation factor 2 subunit 1 (EIF2S1) is among these candidates. EIF2S1 is known as a master regulator of cellular stress responses that additionally regulates circadian rhythms. We show that knockdown of EIF2S1 furthermore impairs temperature compensation, suggesting that the role of CPSF6 in temperature compensation may be mediated by its regulation of EIF2S1.

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Schmal, C., Maier, B., Ashwal-Fluss, R., Bartok, O., Finger, A.-M., Bange, T., Koutsouli, S., Robles, M. S., Kadener, S., Herzel, H., Kramer, A.. 2021-10-08. An integrative omics approach reveals posttranscriptional mechanisms underlying circadian temperature compensation. https://doi.org/10.1101/2021.10.06.463236

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