Physiological Studies on Candida albicans

Candida albicans is a common opportunistic, dimorphic human fungal pathogen. One of its virulence factors is the morphological switch between yeasts and hyphal or pseudohyphal forms, which can invade tissues and cause damage. Our studies focus on factors regulating pseudohyphae and epigenetic modifications of C. albicans. Regulating factors of pseudohyphae are aromatic alcohols and high phosphate. At low concentrations, exogenous aromatic alcohols induced pseudohyphae, as did high phosphate. For addressing the pathways involved in inducing pseudohyphae by aromatic alcohols or high phosphate, we used mutants defective in cAMP dependent PKA pathway (efg1/efg1), MAP kinase pathway (cph1/cph1), or both (cph1/cph1/efg1/efg1). These mutants failed to produce either hyphae or pseudohyphae in the presence of aromatic alcohols; but high phosphate still stimulated pseudohyphae. Gcn4, a transcription activator of more than 500 amino acid related genes, is turned-on in response to amino acid starvation. The accumulation of aromatic alcohols sends nitrogen starvation signals, which inhibit eIF2B, which in turn derepresses Gcn4p. High phosphate also induces pseudohyphae by derepressing Gcn4p, although the pathways involved are still unknown. In sum, aromatic alcohols and high phosphate induce pseudohyphae by derepressing Gcn4. In this study we found a novel posttranslational histone modification in C. albicans, which is biotinylation. Western blot and Mass spectrometry techniques were used to find that Histones H2B and H4 were biotinylated at every condition tested such as yeast vs. hyphae, aerobic growth vs. anaerobic growth, rich medium vs. defined medium. In C. albicans lysines K8, K11 in histone H4 and lysines K17, K18, K31 in histone H2B are biotin attachment sites as found using mass spectrometry. Biotin was also found to enhance the germ tube formation of C. albicans. Germ tube formation assays with biotin-starved cells as inoculum showed low percent of germ tubes (1-5%). Addition of biotin to the media showed 100% germ tubes. Biotinylation of histones were not detected from biotin-starved cells. Appendix-A details work related to Farnesol quantification assays in several strains of C.albicans and Ceratocystis ulmi, and growth studies of class E VPS strains of Saccharomyces Cerevisiae. Adviser: Kenneth W. Nickerson

An Analysis of Anchitherine Equids Across the Eocene–Oligocene Boundary in the White River Group of the Western Great Plains

Anchitherine horses are a subfamily of equids that are abundantly represented in the late Eocene and early Oligocene of North America. This group has been heavily studied in the past, but important questions still remain. Some studies have focused on the Eocene-Oligocene boundary and have used these equids along with other taxa to study mammalian diet and climate change through this interval. I reexamine two anchitherine genera, Mesohippus and Miohippus, from stratigraphic sequences of the White River Group in western Nebraska and southwestern South Dakota. These sequences span the Chadronian (late Eocene), Orellan (early Oligocene), and Whitneyan (early Oligocene) North American land-mammal ages. The most recent revision of these genera was done by Prothero and Shubin (1989). I review the characters used for taxonomic identification. This includes characters such as the hypostyle, the articular facet on the third metatarsal, and dental dimensions. To avoid possible biases caused by combining specimens from different stratigraphic levels, specimens were separated by location and stratigraphic level. The length and width of cheek teeth, and tooth rows were measured on 488 specimens. First molar area serves as a proxy for body mass in horses and other mammals, and can be useful for distinguishing among species. Results indicate that the characters used by Prothero and Shubin were highly variable in anchitherine horses and are not useful for distinguishing between these genera. The development of the articular facet on the third metatarsal may be a function of body size and therefore may be of no more utility than first molar area. Variability in first molar area suggests the presence of three species in the medial and late Chadronian, two species in the Orellan, and at least two species in the Whitneyan. Due to a lack of objective criteria separating Mesohippus from Miohippus, I recommend synonymy of these genera, making Mesohippus a junior subjective synonym.