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Multi-copy suppressor screen for Cryptococcus neofomans RAS1



Multi-copy suppressor screen for Cryptococcus neofomans RAS1



Abstracts of the General Meeting of the American Society for Microbiology 103: F-011



Background: Cryptococcus neofomans is a human fungal pathogen, and its ability to grow at high temperature is a key feature in pathogenesis. Previous results place the small G-protein Ras1 as the upstream factor in a signaling pathway required for high temperature growth for this basidiomycete. The goal of our current work is to identify other components of this pathway that would act downstream of Ras1. Methods/Results: A multi-copy suppressor screen was performed in the C. neofomans ras1 mutant strain. ras1 mutants were transformed with a library of genomic DNA into the telomeric plasmid pPM8. The transformants were screened for restoration of the ras1 mutant high temperature growth defect. Several plasmids were rescued, and the genomic inserts were characterized using available C. neoformans genome databases. Among the putative genes found in the inserts, special attention has been paid to genes encoding a putative Zn-finger protein, a DNA mis-match repair protein, and a Rac1 homolog. Ras proteins in mammalian cell lines activate Rac proteins. Conclusions: A multi-copy suppressor approach has identified putative downstream effectors of the C. neoformans Ras1 high temperature growth pathway.

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Accession: 035349086

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