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Characterization and Role of p53 Family Members in the Symbiont-Induced Morphogenesis of the Euprymna scolopes Light Organ

Michael S. Goodson, Wendy J. Crookes-Goodson, Jennifer R. Kimbell and Margaret J. McFall-Ngai^*

Department of Medical Microbiology and Immunology, University of Wisconsin, Madison, Wisconsin 53706

^* To whom correspondence should be addressed: Department of Medical Microbiology and Immunology, University of Wisconsin, 1300 University Avenue, Madison, Wisconsin 53706. E-mail: mjmcfallngai{at}wisc.edu

Within hours of hatching, the squid Euprymna scolopes forms a specific light organ symbiosis with the marine luminous bacterium Vibrio fischeri. Interactions with the symbiont result in the loss of a complex ciliated epithelium dedicated to promoting colonization of host tissue, and some or all of this loss is due to widespread, symbiont-induced apoptosis. Members of the p53 family, including p53, p63, and p73, are conserved across broad phyletic lines and p63 is thought to be the ancestral gene. These proteins have been shown to induce apoptosis and developmental morphogenesis. In this study, we characterized p63-like transcripts from mRNA isolated from the symbiotic tissues of E. scolopes and described their role in symbiont-induced morphogenesis. Using degenerate RT-PCR and RACE PCR, we identified two p63-like transcripts encoding proteins of 431 and 567 amino acids. These transcripts shared identical nucleotides where they overlapped, suggesting that they are splice variants of the same gene. Immunocytochemistry and Western blots using an antibody specific for E. scolopes suggested that the p53 family members are activated in cells of the symbiont-harvesting structures of the symbiotic light organ. We propose that once the symbiosis is initiated, a symbiont-induced signal activates p53 family members, inducing apoptosis and developmental morphogenesis of the light organ.

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