Cell Division

Errors in chromosome transmission alter the balance of tumour suppressor and tumour promoter genes. This facilitates subsequent changes in genome composition in the ensuing cell divisions, ultimately leading to transformation and cancer. Cell cycle controls ensure that a cell does not segregate its replicated genomes until a number of criteria are fulfilled. Not only must cells have undergone the appropriate degree of growth to justify division and receive the right external cues, but the genome must also be intact and any damage that has occurred since DNA replication must have been repaired. Mitotic checkpoint networks within mitosis then ensure that the splitting of the chromosomes into two identical chromatids does not happen until all of the chromosomes have attached to both spindle poles and cells do not leave mitosis until all stages of mitosis have been done correctly.

Cell cycle control in fission yeast

The molecules that regulate progression through the cell division cycle are essentially the same in all eukaryotes from yeast to man. Yeast are single, free living fungi whose prime objective, in the right conditions, is to grow and divide. They have a short cell division cycle, are genetically malleable, have good cytology and are amenable to study by biochemistry. Studies of progression through the relatively simple yeast cell division cycle have therefore often paved the way for the study of more complex processes in human cells. Many of the molecules that regulate cell division in humans were first identified by studies in simple model systems such as yeast, fruit flies or frog eggs. We study cell division in fission yeast. 

Regulating commitment to mitosis from the spindle pole

Commitment to mitosis is regulated by the activity of a protein kinase complex called MPF. During interphase Wee1 related kinases phosphorylate and inhibit MPF. This phosphate is removed by Cdc25 phosphatases to promote mitotic entry. MPF activation promotes a positive feedback loop that boosts Cdc25 and inhibits Wee1 activity. This feedback loop includes polo kinase. The spindle pole component Cut12 associates with polo kinase, modulates its activity and can be mutated to promote mitosis in cells that lack Cdc25. This suggests that commitment to mitosis is promoted by events on the spindle pole. 

 

Protein Kinases & Mitotic Control

 

MPF activation triggers downstream kinases

Although MPF directly activates a limited number of molecules that mediate chromosome segregation, the principle mechanism by which it promotes mitosis is by activating a set of downstream kinases that includes kinases related to Polo, aurora, NIMA and Mps1. Activation of these kinases promotes the formation of the condensation of chromosomes, the attachment of the chromosomes to the spindle and the formation of the polo, aurora and NIMA kinase.

Identification of novel spindle components

One of the major advantages of yeast for the study of cell processes is the ability to use genetic screens to identify molecules without any prior knowledge of the type, abundance or physical properties of the molecule that is being looked for. The genetic screen simply has to be constructed in a way that it selects mutants in the process of interest from the background of all possible mutations in the genome. We have therefore screened for mutants that are able to form a mitotic spindle at 25°C but not at 36°C. We then clone and study the genes that are mutated in these spindle architecture defective (sad) mutants.