RRC ID 49280
Author Sugiyama M, Sakaue-Sawano A, Iimura T, Fukami K, Kitaguchi T, Kawakami K, Okamoto H, Higashijima S, Miyawaki A.
Title Illuminating cell-cycle progression in the developing zebrafish embryo.
Journal Proc Natl Acad Sci U S A
Abstract By exploiting the cell-cycle-dependent proteolysis of two ubiquitination oscillators, human Cdt1 and geminin, which are the direct substrates of SCF(Skp2) and APC(Cdh1) complexes, respectively, Fucci technique labels mammalian cell nuclei in G(1) and S/G(2)/M phases with different colors. Transgenic mice expressing these G(1) and S/G(2)/M markers offer a powerful means to investigate the coordination of the cell cycle with morphogenetic processes. We attempted to introduce these markers into zebrafish embryos to take advantage of their favorable optical properties. However, although the fundamental mechanisms for cell-cycle control appear to be well conserved among species, the G(1) marker based on the SCF(Skp2)-mediated degradation of human Cdt1 did not work in fish cells, probably because the marker was not ubiquitinated properly by a fish E3 ligase complex. We describe here the generation of a Fucci derivative using zebrafish homologs of Cdt1 and geminin, which provides sweeping views of cell proliferation in whole fish embryos. Remarkably, we discovered two anterior-to-posterior waves of cell-cycle transitions, G(1)/S and M/G(1), in the differentiating notochord. Our study demonstrates the effectiveness of using the Cul4(Ddb1)-mediated Cdt1 degradation pathway common to all metazoans for the development of a G(1) marker that works in the nonmammalian animal model.
Volume 106(49)
Pages 20812-7
Published 2009-12-8
DOI 10.1073/pnas.0906464106
PII 0906464106
PMID 19923430
PMC PMC2779202
MeSH Animals Animals, Genetically Modified Biomarkers / metabolism Cell Cycle* Cell Nucleus Embryo, Nonmammalian / cytology* Embryonic Development* Fluorescent Dyes / metabolism G1 Phase Mitosis Molecular Sequence Data Notochord / cytology Retina / cytology Retina / embryology S Phase Time Factors Zebrafish / embryology* Zebrafish / genetics
IF 9.412
Times Cited 131
DNA material mKO2-zCdt1(1/190) / pT2KXIGdeltain (RDB15275) mAG-zGeminin(1/100) / pT2KXIGdeltain (RDB15276) mAG-hGeminin (1/60) / pT2KXIGdeltain (RDB15277) mCherry-zCdt1(1/190) / pT2KXIGdeltain (RDB15278) mVenus-zGeminin (1/100) / pT2KXIGdeltain (RDB15279)