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The wonderful duplicating “machine” A briefe history of the cell cycle • 18xx Interphase-Mitosis (Flemming) • 1951 Discovery of a distinct S-phase (Howard and Pelc) • 1970 Cell fusion experiments (Rao and Johnson) • 1971 Discovery of MPF (Masui and Markert) • 197x Yeast cdc mutants (Nurse, Hartwell etc.) • 1983 Discovery of the first cyclins (Hunt) • 1988 Cyclin B+cdc2 = MPF (Maller, Newport, Beach) • 18xx Interphase-Mitosis (Flemming) • 1951 Discovery of a distinct S-phase (Howard and Pelc) • 1970 Cell fusion experiments (Rao and Johnson) • 1971 Discovery of MPF (Masui and Markert) • 197x Yeast cdc mutants (Nurse, Hartwell etc.) • 1983 Discovery of the first cyclins (Hunt) • 1988 Cyclin B+cdc2 = MPF (Maller, Newport, Beach) QuickTime™ and a Sorenson Video decompressor are needed to see this picture. QuickTime™ and a Video decompressor are needed to see this picture. • 18xx Interphase-Mitosis (Flemming) • 1951 Discovery of a distinct S-phase (Howard and Pelc) • 1970 Cell fusion experiments (Rao and Johnson) • 1971 Discovery of MPF (Masui and Markert) • 197x Yeast cdc mutants (Nurse, Hartwell etc.) • 1983 Discovery of the first cyclins (Hunt) • 1988 Cyclin B+cdc2 = MPF (Maller, Newport, Beach) Fluorescent Activated Cell Scan/Sorter (FACS) A typical FACS analysis Methods for cell synchronisation • 18xx Interphase-Mitosis (Flemming) • 1951 Discovery of a distinct S-phase () • 1970 Cell fusion experiments (Rao and Johnson) • 1971 Discovery of MPF (Masui and Markert) • 197x Yeast cdc mutants (Nurse, Hartwell etc.) • 1983 Discovery of the first cyclins (Hunt) • 1988 Cyclin B+cdc2 = MPF (Maller, Newport, Beach) Cell fusion experiment Johnson and Rao 1971 G1 M M S M M G2 M M • 18xx Interphase-Mitosis (Flemming) • 1951 Discovery of a distinct S-phase (Howard and Pelc) • 1970 Cell fusion experiments (Rao and Johnson) • 1971 Discovery of MPF (Masui and Markert) • 197x Yeast cdc mutants (Nurse, Hartwell etc.) • 1983 Discovery of the first cyclins (Hunt) • 1988 Cyclin B+cdc2 = MPF (Maller, Newport, Beach) Maturation promoting factor cytostatic factor Discovery of MPF Masui and Markert 1971 Oscillations of MPF • 18xx Interphase-Mitosis (Flemming) • 1951 Discovery of a distinct S-phase (Howard and Pelc) • 1970 Cell fusion experiments (Rao and Johnson) • 1971 Discovery of MPF (Masui and Markert) • 197x Yeast cdc mutants (Nurse, Hartwell etc.) • 1983 Discovery of the first cyclins (Hunt) • 1988 Cyclin B+cdc2 = MPF (Maller, Newport, Beach) Model organisms I - Budding yeast (S.cerevisiae) L. Hartwell Model organisms II - Fission yeast (S.pombe) P. Nurse Generation of temperature sensitive budding yeast mutants Generation of temperature sensitive budding yeast mutants Analysis of ts cell cycle mutants G1 S M M M M Identification of cell cycle ts mutants Some ts mutants which became famous The two models of cell cycle regulation • 18xx Interphase-Mitosis (Flemming) • 1951 Discovery of a distinct S-phase (Howard and Pelc) • 1970 Cell fusion experiments (Rao and Johnson) • 1971 Discovery of MPF (Masui and Markert) • 197x Yeast cdc mutants (Nurse, Hartwell etc.) • 1983 Discovery of the first cyclins (Hunt) • 1988 Cyclin B+cdc2 = MPF (Maller, Newport, Beach) • 18xx Interphase-Mitosis (Flemming) • 1951 Discovery of a distinct S-phase (Howard and Pelc) • 1970 Cell fusion experiments (Rao and Johnson) • 1971 Discovery of MPF (Masui and Markert) • 197x Yeast cdc mutants (Nurse, Hartwell etc.) • 1983 Discovery of the first cyclins (Hunt) • 1988 Cyclin B+cdc2 = MPF (Maller, Newport, Beach) Oscillations of mitotic cyclins are required for MPF activity Regulation of mitotic cyclin levels in cycling cells Cdc2/28 (=cdk1) is regulated by: 1. Binding of a cyclin 2. Inhibitory phosphorylation 3. Activating phosophorylation 4. Binding of CKI 5. Cellular localization QuickTime™ and a Animation decompressor are needed to see this picture.