1. Expression of neuronal protein Tau in
Candida albicans, strain 36232
College of Staten Island
Nicholas Ferraro
2. Overview of Candida albicans
• Dimorphic fungal pathogen.
• Budding form and filamentous form which contributes to host tissue invasiveness and fungal
pathogenicity.
• Cells adhere during hyphae formation resulting in biofilm formation which is a polymeric
conglomeration of extracellular proteins and polysaccharides.
• Hyphae formation is mediated by actin and microtubules.
• Actin plays a significant role in elongation of filaments.
• Microtubules function in nuclei positioning in filaments and regulation of hyphael
morphogenesis.
3. Neuronal protein Tau
• Tau is a hydrophilic phosphoprotein.
• Tau plays a role in microtubule assembly, stabilization and bundling in the neurons of the mammalian
brain.
• Phosphorylation of Tau is developmentally regulated and the level of Tau phosphorylation controls
the normal interactions with microtubules.
• When Tau is in a hyperphosphorylated state, it dissociates from microtubules and self assemblies into
filaments and neurofibrillary tangles, as seen in Alzheimer's disease.
10. Biofilm Assay
• In well “A” containg the parental
strain 36232 we can see strong
adherence compared with 36232-
CMV and 36232-CMV-Tau. This
is indicating that the more
adherence to the well the less
filamentation occurring.
11. C. Albicans 36232 C.Albicans 36232-CMV C. Albicans 36232-CMV -Tau
24 hour incubation
22. Tau expression in: 36232, 36232-CMV, and 36232-CMV-Tau
• At 24 hours we see the first signs of filamentation occurring in the CMV derivative. Few filaments are seen in
36232 with non in 36232-CMV-Tau.
• At 48, and 72 hours we begin to see long filaments in every field of 36232-CMV as well as short filaments
with adhesion in 36232-CMV-Tau. There is still few filamentous growth in 36232.
• At 96 hours we see large long filaments with in 36232, and long filaments in 36232-CMV. 36232-CMV-Tau
shows short filaments with adhesion.
• At the 7 day marker, we see similar filamentation in both 36232 and 36232-CMV. In 36232-CMV-Tau we still
see small aggregates of short filaments with much adhesion.
• With regard to the images obtained from the confocal microscopes we can see labeling of tau in the Tau2
strain however, there is labeling also found in the parental strain.
23. Does Tau effect filamentation?
• In the 7 day period we can see morphological differences in filamentation between 36232, 36232-CMV, and
36232-CMV-Tau. We can see by the seventh day of incubation both 36232 and 36232-CMV-Tau look similar in
filamenation length and adhesion. It is 36232-CMV-Tau that has the significant difference of short filaments with
much adhesion.
• With regard to the images obtained by the confocal microscope, we can see that tau does have an effect on
filamentation however, labeling of both the parental strain and the Tau2 strain indicate that there maybe be cross
interaction between the antibody and another site of homology or protocol revision may be in order.
• Unfortunately, results from the 36232-CMV plasmid control were unable to be obtained. Further analysis such as
Western Blot analysis, NMR and TEM is required to further establish the expression of Tau.
