Pan1 And Genetic Domain Structures

Cin1 exists as Cin1-S (short-form) and Cin1-L (long-form) isoforms as a result of alternative splicing of intron #6. Cin1-S differs from Cin1-L by lacking the C-terminal RhoGEF (DH)-PH domain. We previously generated a CIN1-S mutant strain by replacing the C-terminus with an NAT marker gene. Significantly, we have found that Cin1-S confers a survival advantage over WT in the mouse brain via co-infection. Further investigation will include constructing a mutant expressing the long isoform Cin1-L. We have generated a partial CIN1-L point-mutation allele linked to G418 resistance, but have yet to obtain the Cin1-L mutant through the conventional gene replacement method. Recently, we have developed a one-step CRISPR-CAS9 technique for mutagenesis in Cn. In this aim, we will generate the Cin1-L mutant, characterize its function, and perform co-infection assays. Moreover, we will perform dual RNA-Seq to profile whether there is differential gene expression in infection by Cin1-S and Cin1-L in order to address the mechanism associated with Cin1-S CNS survival

It is well-known that physiological differences exist between pathogens grown during in vitro versus in vivo conditions. In Cn, differences exist in the production of virulence factors such as melanin and capsule in the laboratory under nutrient-rich conditions compared to tissue culture medium and in the host. Furthermore, these physiological differences affect the organism’s sensitivity to antifungal drugs, and are also likely to affect production of biomarkers such as exRNAs [29]. A significant body of data is emerging that indicates an abundance of (EVs)-exosomes containing exRNA

Several proteins have been thought to have a part in regulating AURA activity and/or expression, such as CHFR. There are no previous reports on the localization of this protein to primary cilia, but it is believed that CHFR may function at the cilium to encourage cilia stability through inactivation of AURA. Knockdown of AURA by siRNA has also led to the proposal that tubulin deacetylase(s) is not the main target for AURA-induced ciliary disassembly or inhibition of ciliary assembly in OSE

DCM is still the leading cause of morbidity and mortality between diabetic patients. The complete understanding of DCM pathophysiology and the role of exosomes in this pathologic disorder are still being investigated. However, there has been emerging in vitro ex-vivo and in vivo experimental data about different underlying mechanisms that might be helpful to answer important questions raised in the study of DCM. Over the last decade, exosomes have been extensively studied as microRNA-carriers which are pivotal for local tissue-specific and inter-systemic cells communication. Of interest, miRNAs-enriched exosomes play an important role in development of DCM based on accumulated evidences such as alteration of miRNAs profile in the diabetic heart,

The cytochalasins are cell-permeable fungal metabolites inhibiting actin polymerization. This interferes with such diverse processes as cell growth, movement, phagocytosis, degranulation, as well as secretion.

Cellular Basis Of Disease: Why has the discovery of Toll-like receptors revolutionised our understanding of how the innate immune system works, and what is the therapeutic potential?

Many individuals in today’s world don’t know what to think of when someone says the word yeast, even though it’s all around us. What is it? What does it do and how does it do what it does? For starters, yeast is a unicellular eukaryote that is a member of the fungus family. Its main function and purpose is to perform aerobic respiration and anaerobic respiration, otherwise known as fermentation. It is through these processes that yeast produces carbon dioxide, ethanol, lactate, and water, which humans utilize in the production of bread, alcohol, and medicines.

The surface projections on HCL cells have multiple functions. The fine hair-like cytoplasmic structures on the surface of these cells are rich in F actin (polymerized actin). Through constitutive activation of Rho-GTPases, and overexpression of proteins, such as CD9, pp52, and GAS7 (growth arrest–specific 7), the F-actin filaments crosslink and form aggregates (Burthem, et al., 1994). These aggregated structures are

At present, there are no experimental models allowing direct interactions between Cn and neurons. Macrophages, on the other hand, play an important role in the early immune response against Cn and a macrophage-like J774A.1 cell line model is available for studying the Cn-macrophage interaction [51]. When appropriately stimulated, macrophage-like J774A.1 cells exhibit measurable parameters in uptake, ingestion, and clearance or lysis of fungal cells that correlate with in vivo infection outcomes. We have previously employed the J774A.1 cell line to successfully examine gene expression following phagocytosis of the crg2 mutant lacking a regulator of G protein signaling protein homolog, Crg2 [51].

exRNA was submitted to the Beijing Genome Institute (BGI, Shenzhen, China) for cDNA synthesis and RNA-Seq. Approximately 100 ng cDNA from each strain was each subject to illumina deep sequencing using BGISEQ-500 technology. A total of 27,084,174 clean sRNA reads was obtained from the cin1 mutant and 27,402,307 clean reads were obtained from WT (Table 1). The length of sRNA is between 18 nt and 30 nt. The initial sRNA annotation provided by BGI suggested that approximately 96% sRNA sequences were mapped to the available sRNA database including miRBase and Rfam. Further annotation, prediction, expression, screening of differentially expressed sRNA, and miRNA target prediction will be performed under the direction of Dr. Taylor. Using this methodology, we will determine the role of Cin1 in secretion and uptake of sRNA.

It was also reported that the MATα strain could sustain dominance over the isogenic MATa strain in the brain, raising an interesting aspect of virulence while substantiating the result of an early report {Nielsen, 2005 #5284; Kwon-Chung, 1992}. The intriguing finding that Cin1-S maintains a CNS survival advantage provides a novel avenue to probe the CNS propensity of Cn. Finally, given the importance of exRNA in cellular communications, it is tempting to speculate that Cin1 regulates uptake and export of exRNA to play a role in host-pathogen interactions and pathogenesis. Taken together, we may possess an unprecedented system to probe the mechanisms of pathogenesis in Cn.

Aim 1 is to select and validate the expression of candidate ex-RNAs produced by Cn in vitro.

Overexpression of the HEPN domain in HeLa cells following three hours of starvation indicates that sacsin plays a key role in lysosomal transport due to the reduced efficiency of perinuclear lysosomal clustering. HEPN’s property of dimerizing may be disrupting the function of with full-length sacsin by preventing endogenous sacsin from dimerizing within the cell. Thus, dimerization may be essential to sacsin’s function, particularly for binding JIP3, which is necessary for lysosomal transport7. HATPase 3 overexpression may be occupying JIP3 binding, but was not found to disrupt lysosomal localization following starvation, perhaps due to the truncated size of this deletion construct.

The goal of this proposed study is to test the hypothesis that replicative life span (RLS) in Candida glabrata enhances azole resistance through regulation of efflux pumps that subsequently leads to drug resistance during infection. Aim 1 is to test whether aging-mediated azole resistance is the result of enhanced transcription for major drug pumps or transport facilitors. Aim 2 is to determine whether the enhanced antiazole activity of the older cells contribute to "clinical" drug resistance. Establishing a correlation between RLS and pathogenicity is a novel concept that the PI has successfully established for the fungal pathogen, Cryptococcus neoformans. In addition, the PI has powerful tools such as the HYAA chip and is poised to

Saccharomyces cerevisiae is widely recognized as an ideal eukaryotic organism for biochemical and genetic studies. Although yeasts have greater genetic complexity than bacteria, they still share many of the technical advantages that permitted rapid progress in the molecular genetics of eukaryotes and their viruses. Some of the properties that make yeast suitable for studies include the existence of both stable haploids and diploids, rapid growth, clonability, and the ease of replica plating and mutant isolation.