Fluorescence fluctuation analysis reveals PpV dependent Cdc25 protein dynamics in living embryos


Autoři: Boyang Liu aff001;  Ingo Gregor aff003;  H.-Arno Müller aff004;  Jörg Großhans aff001
Působiště autorů: Fachbereich Biologie (FB17), Philipps-Universität Marburg, Marburg, Germany aff001;  Institut für Entwicklungsbiochemie, Universitätsmedizin, Georg-August-Universität Göttingen, Göttingen, Germany aff002;  Drittes Physikalisches Institut, Georg-August-Universität Göttingen, Göttingen, Germany aff003;  Fachgebiet Entwicklungsgenetik, Institut für Biologie, Universität Kassel, Kassel, Germany aff004
Vyšlo v časopise: Fluorescence fluctuation analysis reveals PpV dependent Cdc25 protein dynamics in living embryos. PLoS Genet 16(4): e32767. doi:10.1371/journal.pgen.1008735
Kategorie: Research Article
doi: 10.1371/journal.pgen.1008735

Souhrn

The protein phosphatase Cdc25 is a key regulator of the cell cycle by activating Cdk-cyclin complexes. Cdc25 is regulated by its expression levels and post-translational mechanisms. In early Drosophila embryogenesis, Cdc25/Twine drives the fast and synchronous nuclear cycles. A pause in the cell cycle and the remodeling to a more generic cell cycle mode with a gap phase are determined by Twine inactivation and destruction in early interphase 14, in response to zygotic genome activation. Although the pseudokinase Tribbles contributes to the timely degradation of Twine, Twine levels are controlled by additional yet unknown post-translational mechanisms. Here, we apply a non-invasive method based on fluorescence fluctuation analysis (FFA) to record the absolute concentration profiles of Twine with minute-scale resolution in single living embryos. Employing this assay, we found that Protein phosphatase V (PpV), the homologue of the catalytic subunit of human PP6, ensures appropriately low Twine protein levels at the onset of interphase 14. PpV controls directly or indirectly the phosphorylation of Twine at multiple serine and threonine residues as revealed by phosphosite mapping. Mutational analysis confirmed that these sites are involved in control of Twine protein dynamics, and cell cycle remodeling is delayed in a fraction of the phosphosite mutant embryos. Our data reveal a novel mechanism for control of Twine protein levels and their significance for embryonic cell cycle remodeling.

Klíčová slova:

Cell cycle and cell division – Drosophila melanogaster – Embryos – Invertebrate genomics – Mitosis – Phosphatases – Phosphorylation – Fluctuation analysis


Zdroje

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