Candida albicans wild type vs eed1 during RHE infection
ABSTRACT: Candida albicans wild type SC5314 and the eed1 delta mutant were used to infect reconstituted human oral epithelium (RHE) for 24h at 37°C and 5% CO2. Samples were taken 1h, 12h and 24h after infection. Total RNA was isolated, labeled with Cy5 and hybridised with a Cy3- labeled common reference.
The extension of germ tubes into elongated hyphae by Candida albicans is essential for damage of host cells. The C. albicans-specific gene EED1 plays a crucial role in this extension and maintenance of filamentous growth. eed1Δ cells failed to extend germ tubes into long filaments and switched back to yeast growth after 3 h of incubation during growth on plastic surfaces. Expression of EED1 is regulated by the transcription factor Efg1 and ectopic overexpression of EED1 restored filamentation in ...[more]
Project description:C. albicans wild type strain SC5314, the eed1 deletion mutant and an eed1 delta mutant overexpressing UME6 (eed1 + pTET-UME6) were grown on plastic (37°C, RPMI1640 medium, plastic surface, 5% CO2) for 12h. Total RNA was isolated using a phenol-chloroform protocol and labeled with Cy5. Cy5- labeled sample RNA was hybridised with Cy3- labeled common reference (SC5314, 37°C, exponential culture).
Project description:The presence of Set2-mediated methylation of H3K36 (K36me) correlates with transcription frequency throughout the yeast genome. K36me targets the Rpd3S complex to deacetylate transcribed regions and suppress cryptic transcription initiation at certain genes. Here, using a genome-wide approach, we report that the Set2-Rpd3S pathway is generally required for controlling acetylation at coding regions. When using acetylation as a functional read-out for this pathway, we discovered that longer genes and, surprisingly, genes transcribed at lower frequency exhibit a stronger dependency. Moreover, a systematic screen using high resolution tiling microarrays allowed us to identify a group of genes that rely on Set2-Rpd3S to suppress spurious transcripts. Interestingly, most of these genes are within the group that depend the same pathway to maintain a hypo-acetylated state at coding regions. These data highlight the importance of using the functional readout of histone codes to define the roles of specific pathways.
Project description:We have established Drosophila melanogaster as a model system for ocular hypertension by expressing wild-type human myocilin (MYOC) in the Drosophila eye. Here, we have created transgenic flies that express four clinically relevant mutant forms of MYOC (R342K, Q368X, D380N and K423E) in their eyes using the gmr-Gal4/UAS binary system. We compare and identify human glaucoma candidate genes based on the transcription profiles of flies that express wt-MYOC or mutant-MYOCs.