ROLE OF THE ANTIZYME FAMILY DURING XENOPUS DEVELOPMENT

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This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. Polyamines are important organic cations that are essential for life. Cells that are actively proliferating such as embryonic cells or cancer cells, have strict needs for polyamines. Thus, manipulation of polyamine levels can have profound effects on growth. Cells maintain well-regulated pathways that control polyamine levels at optimum levels for cell growth. Antizyme (AZ) is a crucial protein that influences the rate of cell proliferation by controlling the levels of polyamines. An aim of this project is to characterize the regulation of polyamine levels and effects on cell growth using a developmental model of proliferation, Xenopus laevis embryogenesis. A focus of the work will be to determine how AZ regulates the proliferative capacity of Xenopus embryonic cells. There has not been a systematic study of antizyme function using a developmental model system such as the amphibian Xenopus laevis. This project will characterize the expression pattern of the AZ gene family using quantitative RT-PCR, Northerns, Westerns, and two dimensional protein gels during the early development of Xenopus embryos. The function of the AZ family will be studied by microinjection experiments that will force AZ expression at the one cell embryo stage. This will allow for a study of whether AZ can dominantly control cell proliferation. Thus, the outcome of this project will be the elucidation of the role of the AZ family in proliferation and development using a tractable organism, Xenopus laevis. Since the developmental pathways of Xenopus laevis are closely related to the same mammalian pathways, this project will help to shed light on the role of AZ in mice and humans with potential applications to the growth of cancer cells.