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Specific roles played by the polyamines in DNA replication

Background

DNA replication is a complex process involving a variety of proteins and other molecules that are required to maintain chromatin in a replication prone state as well as enzymes with their accessory proteins that are required for the incorporation of the nucleotides. DNA replication is impaired in cells depleted of their polyamines and we have evidence indicating that Okazaki fragment maturation is impaired in polyamine-depleted cells.

Experimental system

Mammalian cells seeded in the absence or presence of polyamine biosynthesis inhibitors will be used.

Roles of polyamines in DNA replication

We are investigating effects of polyamine biosynthesis inhibition on the replication machinery. We are presently investigating the rate of DNA replication using DNA fiber autoradiography after labelling cells with bromodeoxyuridine. We will also study the in situ distribution of various proteins involved in DNA replication and mainly Okazaki fragment maturation using confocal laser scanning microscopy after staining cells with fluorescently-labelled specific antibodies.

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