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By Richard Twyman

This booklet emphasises the unifying rules and mechanisms of molecular biology, with common use of tables and packing containers to summarise experimental information and gene and protein services.

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Advanced molecular biology: a concise reference

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Pombe, and similar components are found in other eukaryotes. Essentially, mitosis is initiated bya burst of Cdc2 activity generated by a posttranslational positive feedback mechanism(this differs fromentry into the S phase in S. cerevisiae, which is governed by a transcriptional feedback loop). The mechanism is broadly conserved throughout the eukaryotes, but the checkpoints forentry into mitosis are regulated in different ways, as discussed below. In S. 5). It is activated by dephosphorylation, carried out by Cdc25 and another phosphatase termed Pyp3, and inactivated by redundant kinases termed Weel and Mikl.

Stuwing cell cycle regulation. Two complementary approaches have been used to characterize and isolate the regulatory components of the cell cycle. Inthe heterokaryon approach, nuclei at different stages of the chromosome cycle are joined in a common cytoplasm and their behavior observed. Cultured mammalian cells and amphibian eggs have been used for these experiments. 2. 2 Heterokaryon experiments to investigate regulatoryfactors controlling the cell cycle Fusion Result replicate nuclei Both S X G1 S X G2 M x G,, S or G2 G1 Dresent v x G2 are mitosis S-phase cellcompletesreplication, Gp-phase nucleus waits for S-phase nucleus to complete replication and then both cells enter the M phase Interphase nucleus enters precocious mitosis (regardless of state of chromosome replication) Neither nucleus undergoes replication Both or Conclusion S nucleus contains an S-phase promoting factor G2 nucleuscannotrespond to S-phase activator (a re-replication block), S-phase activator is also an inhibitor of mitosis M nucleus contains an M-phase promoting factor S-phase and M-phase activators 26 Advanced Molecular Biology transplantation studies -interphase nuclei formed spindles when injected into eggs arrested at the metaphase of meiosis I, and cytoplasmfrom these eggs could inducemeiosis in oocytes arrested at G2.

The model proposes that a cytoplasmic licensing factor interacts with DNA origins before reassembly of the nuclear envelope at the telophase. This factor is a target for regulation by S-phasepromoting CDK-cyclin complexes and initiates replication at each origin at the beginning of the S phase, becoming inactivein the process. During the remainder of the cell cycle, the new licensing factor is unable to enter the nucleus, and the precocious initiation of replication is prevented. The factor enters the nucleus at the start of mitosis, but is inactive or prevented from interacting with DNA because of the condensed state of the chromatin.

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