title: Imaging analysis of cell cycle-dependent degradation of Cdt1 in mammalian cells.
abstract: Numerous cell cycle-regulating proteins are controlled by protein degradation. Recent work shows that ubiquitination-dependent proteolysis plays an important role in once-per-cell cycle control of DNA replication. Cdt1 is a licensing factor essential for assembling the pre-replicative complex on replication origins. Cdt1 is present in G1 phase, but after S phase ubiquitin-mediated proteolysis maintains Cdt1 at low levels. This is important to prevent the re-replication of chromosomal DNA. The cell cycle-dependent degradation of Cdt1 can be monitored by dual staining of the cell nuclei with antibodies against Cdt1- and S/G2-phase marker proteins, such as cyclin A or geminin.
authors.affiliation: Graduate School of Life Science, University of Hyogo, Kamigori, Ako-gun, Hyogo, 678-1297, Japan.
authors.forename: Yasushi
authors.initials: Y
authors.lastname: Shiomi
authors.forename: Naohiro
authors.initials: N
authors.lastname: Suenaga
authors.forename: Miyuki
authors.initials: M
authors.lastname: Tanaka
authors.forename: Akiyo
authors.initials: A
authors.lastname: Hayashi
authors.forename: Hideo
authors.initials: H
authors.lastname: Nishitani
mesh_terms: D000906:Antibodies / Q000032:analysis; D002453:Cell Cycle; D018797:Cell Cycle Proteins / Q000032:analysis* / Q000378:metabolism*; D019925:Cyclin A / Q000032:analysis / Q000378:metabolism; D064248:Geminin / Q000032:analysis / Q000378:metabolism; D006367:HeLa Cells; D006801:Humans; D008856:Microscopy, Fluorescence / Q000379:methods*; D061848:Optical Imaging / Q000379:methods; D059748:Proteolysis; D025801:Ubiquitin / Q000378:metabolism
chemical_list: D000906:Antibodies; C467199:CDT1 protein, human; D018797:Cell Cycle Proteins; D019925:Cyclin A; D064248:Geminin; D025801:Ubiquitin