Does mPER2 protein oscillate without its coding mRNA cycling?: post-transcriptional regulation by cell clock

Genes to Cells : Devoted to Molecular & Cellular Mechanisms
Yoshito FujimotoHitoshi Okamura

Abstract

Does the mammalian oscillatory protein mPER2 show the rhythm without its coding mRNA cycling? Here we answer this question by inserting a single copy of exogenous mPer2 gene to a NIH3T3 fibroblasts cell line, using Flp-In system. We generated the stable cell lines which constantly express mRNAs coding either N-terminal FLAG-tagged full length mPER2 (FLAG-mPER2(full)) or its C-terminal deleted form (FLAG-mPER2(1-1068)), which lacks the binding site to mCRY proteins, under the control of human EF-1alpha promoter. Although serum shock induced the rhythm of endogenous clock machinery in these cell lines, it did not initiate the rhythm of exogenously inserted FLAG-mPer2 genes at the mRNA level. In contrast, FLAG-mPER2(full) proteins showed the rhythm without their coding mRNA cycling. Since cells expressing FLAG-mPER2(1-1068) also showed the rhythm of FLAG-mPER2(1-1068) proteins, the direct binding of mCRY and mPER2 seems not necessary for this protein oscillation. This system clearly demonstrates that the intracellular endogenous clock system has an ability to modify the mPer2 gene post-transcriptionally to make mPER2 proteins oscillate without its coding mRNA cycling.

References

Jun 10, 1998·Genes to Cells : Devoted to Molecular & Cellular Mechanisms·T TakumiH Okamura
Aug 26, 1998·The EMBO Journal·T TakumiH Okamura
Jun 10, 2000·Molecular and Cellular Biology·S YamaguchiH Okamura
Aug 23, 2003·Science·Takuya MatsuoHitoshi Okamura
Feb 17, 2005·Molecular and Cellular Biology·Yoshinobu YamamotoHitoshi Okamura

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Citations

Jan 25, 2007·Nature Reviews. Molecular Cell Biology·Monica Gallego, David M Virshup
May 11, 2010·Reproduction, Fertility, and Development·Christine K RatajczakLouis J Muglia
Apr 19, 2008·Cold Spring Harbor Symposia on Quantitative Biology·U AlbrechtJ Ripperger
Apr 19, 2008·Cold Spring Harbor Symposia on Quantitative Biology·E Garbarino-Pico, C B Green
Apr 19, 2008·Cold Spring Harbor Symposia on Quantitative Biology·H Okamura
Apr 19, 2008·Cold Spring Harbor Symposia on Quantitative Biology·K Vanselow, A Kramer
Jan 19, 2011·Journal of Cell Science·Shihoko KojimaCarla B Green
Oct 6, 2010·The Tohoku Journal of Experimental Medicine·Tada-aki KudoHaruhide Hayashi
May 9, 2012·Genetics Research International·Chengwei LiXiaodong Li
Mar 8, 2011·FEBS Letters·Silke Reischl, Achim Kramer
Jan 27, 2016·Molecules and Cells·Marrit Putker, John Stuart O'Neill
Jun 13, 2013·Current Opinion in Neurobiology·Paul Franken
Oct 19, 2017·Cold Spring Harbor Perspectives in Biology·Koji L Ode, Hiroki R Ueda
Oct 3, 2019·Physical Review. E·Paula S Nieto, C A Condat
Jan 26, 2021·The EMBO Journal·Marrit PutkerJohn S O'Neill

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