No description available.

Inclusion body myositis and Alzheimer's disease are age-related disorders characterized in part by the appearance of intracellular lesions composed of filamentous aggregates of the microtubule-associated protein tau. Abnormal tau phosphorylation accompanies tau aggregation and may be an upstream pathological event in both diseases. Enzymes implicated in tau hyperphosphorylation in Alzheimer's disease include members of the casein kinase 1 family of phosphotransferases, a group of structurally related protein kinases that frequently function in tandem with the ubiquitin modification system. To determine whether casein kinase 1 isoforms associate with degenerating muscle fibers of inclusion body myositis, muscle biopsy sections isolated from sporadic disease cases were subjected to double-label fluorescence immunohistochemistry using selective anti-casein kinase 1 and anti-phospho-tau antibodies. Results showed that the alpha isoform of casein kinase 1, but not the delta or epsilon isoforms, stained degenerating muscle fibers in all eight inclusion body myositis cases examined. Staining was almost exclusively localized to phospho-tau-bearing inclusions. These findings, which extend the molecular similarities between inclusion body myositis muscle and Alzheimer's disease brain, implicate casein kinase 1 alpha as one of the phosphotransferases potentially involved in tau hyperphosphorylation.

Migraine is a common, episodic neurological disorder that includes headache, nausea and hypersensitivity to sensory stimuli. During the headache phase of migraine, migraine patients can be especially hypersensitive to thermal stimuli. The unpredictable and episodic nature of migraine makes it difficult to treat and much of the mechanism of migraine has yet to be elucidated. A T44A substitution in casein kinase 1δ is inherited with migraine with aura. A transgenic mouse model suggests that animals with this mutation exhibit increased sensitivity to thermal stimuli after injection with nitroglycerin (NTG). We performed behavior assays that measure animal responses to thermal stimuli, after injection with NTG, a known migraine-inducer in human migraine patients. Female animals with the CK1δ-T44A mutation are more sensitive than wildtype littermates, suggesting a sex difference emerges in pain sensitivity in animals that express the CK1δ-T44A but not in wildtype siblings. Female CK1δ-T44A animals are more sensitive to the effects of NTG on pain than male CK1δ-T44A mice. This indicates a potential sex hormone related pain response. Since estrogen is implicated in both migraine and pain response, we test the thermal sensitivity of heterozygous ERβKO/+ and CK1δ-T44A; ERβKO/+ mice compared to wildtype and CK1δ-T44A mice. Overall thermal sensitivity is decreased before stress of injection in both male and female ERβKO/+ and CK1δ-T44A: ERβKO/+ mice. This demonstrates that ERβ is necessary for thermal nociception in untreated mice. However, after injection with saline or NTG, animals of all genotypes responded to thermal stimuli similarly. This suggests that estrogen signaling through ERβ is likely not part of the pathway of NTG-induced thermal sensitivity or that one copy of ERβ is sufficient for NTG-induced thermal sensitivity. Since ERβ is fully functional in CK1δ-T44A mice and CK1δ-T44A mice have wildtype thermal sensitivity at baseline, we can conclude that CK1δ-T44A does not modulate ERβ to affect thermal sensitivity in untreated animals.

No description available.

Rift Valley fever virus (RVFV) is a negative-sense RNA virus that is classified as an overlap select agent by the USDA and the HHS. It was first discovered in the Rift Valley of Kenya in the early 1930s. RVFV is an arbovirus that is transmitted by mosquitoes and infects ruminants and humans. RVFV in humans causes an acute self-limiting febrile illness but in a small percentage of cases, a severe version is noted by ocular disease, hepatitis, hemorrhagic fever, and death. In ruminants, the disease is similar with young livestock being the most susceptible. RVFV is also known to cause "abortion storms" where infected pregnant ruminants abort their fetuses with a near 100% fatality rate. Viruses are obligate intracellular parasites utilizing host-factors to replicate. This study identified three host-protein interactors of the viral Gn and L proteins that aid in viral replication. UBR4 was determined to be an interactor of Gn via immunoprecipitation followed by either LC/MS/MS or western blot analysis. Its inhibition via siRNA or CRISPR-Cas9 knockout showed a reduction of viral titers and viral RNA production. It was determined that UBR4 specifically affects viral RNA production and not entry or egress. Conversely, CK1α and PP1α were identified as binding partners of the L protein using similar methods. CK1α, a kinase, and PP1α, a phosphatase, were chosen for further verification due to data demonstrating the L protein is phosphorylated on at least one serine residue, in addition to PP1α already being shown to impact RVFV replication. Inhibition of CK1 and PP1 via small molecule inhibitors, D4476 and 1E7-03, respectively, showed a decrease in viral titers and RNA production. Strand-specific RT-qPCR demonstrates that CK1 and PP1 impact genomic replication. Upon treatment with D4476 a decrease in L protein phosphorylation was observed. Additionally, it has already been shown that treatment with 1E7-03 increases L protein phosphorylation. These data indicate that CK1 and PP1 modulate L protein phosphorylation, contributing to changes in RVFV replication. This study identifies three host-proteins that affect viral replication, which could be used as a foundation for host-based therapeutic and vaccine development. / Doctor of Philosophy / Rift Valley fever virus (RVFV) is a major biological threat due to its ability to infect both livestock and humans and be passed by mosquito bite. RVFV was first discovered in Africa in the early 1930s. To date, there is no approved therapeutic or vaccine. RVFV usually causes very mild disease but in a small percentage of cases, it progresses to include liver disease, vision loss, swelling of the brain, bleeding, and death. A virus itself is not alive; it needs a living host in order to replicate. To do this, it utilizes things naturally occurring inside the host. The purpose of this study is to identify host-factors that the virus uses in order to efficiently make more viruses. The first viral protein of interest is the glycoprotein, Gn, which is important for viral entry and assembly of the viral particles. It was determined that the host-protein UBR4 is an interactor of Gn and that the inhibition of UBR4 decreases the amount of infectious virus being produced. Similarly, the host-proteins, CK1α and PP1α, were found to be interactors of the viral L protein. The L protein is responsible for synthesizing the building blocks of the virus. It was determined that when CK1 and PP1 are inhibited, the L protein is less efficient at making these building blocks. Understanding the host-factors the virus utilizes is important to the basic understanding of how RVFV infects the host and the development of therapeutics to combat an outbreak.