Structure Analysis and Proton Assignment of Mucin Derivative Peptides with Unnatural Amino Acids on the Backbone by NMR Spectroscopy

Color poster with text, charts, and images.The purpose of this project was to acquire two-dimensional proton NMR data for two synthesized mutant mucin peptides, make assignments for all the hydrogen atoms on the peptides, and analyze for any unique structure present. Mucin peptides are considered to have potential applications in the area of cancer vaccine according to many studies on their biological activity. Two short mucin peptide derivatives based on the native sequence Gly-Val-Thr-Ser-Ala-Pro-Asp were synthesized with the Pro residue substituted by cyclohexylcarboxylic acid (cyHCA) and 4-aminobenzoate (4-aBz). Two-dimensional proton NMR data were used to make assignments of all the hydrogen atoms on the peptides and to verify that the synthesized peptides contained the intended sequences. Hydrogen atoms on the peptides that are in close spatial proximity were analyzed for unique structural features of the peptides. The NMR data collected for each peptide correlate with the expected data based on the known structure of the mucin peptide.University of Wisconsin--Eau Claire Office of Research and Sponsored Program

Zebrafish, Genes, and Human Kidneys : Gene Mapping in Zebrafish Mutants May Help Uncover Genetic Roots of Polycystic Kidney Disease

Color poster with text, images, charts, and photographs.Patients with autosomal dominant polycystic kidney disease (ADPKD) suffer from disrupted kidney function due to large, fluid-filled cysts that form in the collecting ducts and kidney tubules. Current treatments for the disease manage cyst growth but do not prevent cyst formation. Although the specific genes that underlie polycystic kidney disease have been identified, the intervening steps between the altered gene and the disease symptoms remain in completely defined. The Lyman Gingerich lab studies polycystic kidney disease using the model organism Danio rerio, the zebrafish. Spinner mutant zebrafish develop kidney cysts and may be a good model for understanding cystic kidney disease.University of Wisconsin--Eau Claire Office of Research and Sponsored Program

Worms, Genetics and Healthy Kidneys : the Candidate PKD-2 Localization Factor, papl, May Play a Role in Polycystic Kidney Disease

Color poster with text, images, charts, and diagrams.Polycystic Kidney Disease (PKD) causes cysts to form within the kidneys, leading ultimately to renal failure. Prior research in our lab demonstrated that expression of the papl gene is downregulated in a zebrafish model of cystic kidney disease. We asked whether the papl gene has a direct effect on proper localization of PKD2, one of the proteins known to be involved in cyst formation, and whether the papl gene influences cilia structure. Neither the human ortholog (ACP7) nor the zebrafish ortholog (acp7/papl-1) has been associated with a specific function or organelle. papl encodes an acid phosphatase with putative hydrolase activity and a metal ion binding domain. Acid phosphatases have been associated with several human disorders, including prostate cancer. Zebrafish papl ESTs have been identified in the kidney, olfactory rosettes, and reproductive system.University of Wisconsin--Eau Claire Office of Research and Sponsored Program

Laying the Groundwork for Investigating the Link Between Genes and Polycystic Kidney Disease

Color poster with text, images, charts, photographs and graphs.Our lab studies human polycystic kidney disease using zebrafish and C. elegans as models. Patients with autosomal dominant polycystic kidney disease (ADPKD) suffer from disrupted kidney function due to large, fluid-filled cysts that form in the collecting ducts and kidney tubules. Current treatments for the disease manage cyst growth but do not prevent cyst formation. Although the specific mutations that underlie polycystic kidney disease have been identified, the intervening steps between the altered gene and the disease symptoms remain unclear. We are particularly interested in the link between primary cilia and kidney cysts. This year, we worked to amplify target zebrafish genes, maintain healthy zebrafish and nematode populations, and study the effects in C. elegans of downregulating cilia-related genes through RNA interference (RNAi).University of Wisconsin--Eau Claire Office of Research and Sponsored Program

TRIM33 Is a Co-Regulator of Estrogen Receptor Alpha

Estrogen receptor alpha (ER)-positive breast cancer is responsible for over 60% of breast cancer cases in the U.S. Among patients diagnosed with early-stage ER+ disease, 1/3 will experience recurrence despite treatment with adjuvant endocrine therapy. ER is a nuclear hormone receptor responsible for estrogen-driven tumor growth. ER transcriptional activity is modulated by interactions with coregulators. Dysregulation of the levels of these coregulators is involved in the development of endocrine resistance. To identify ER interactors that modulate transcriptional activity in breast cancer, we utilized biotin ligase proximity profiling of ER interactomes. Mass spectrometry analysis revealed tripartite motif containing 33 (TRIM33) as an estrogen-dependent interactor of ER. shRNA knockdown showed that TRIM33 promoted ER transcriptional activity and estrogen-induced cell growth. Despite its known role as an E3 ubiquitin ligase, TRIM33 increased the stability of endogenous ER in breast cancer cells. TRIM33 offers a novel target for inhibiting estrogen-induced cancer cell growth, particularly in cases of endocrine resistance driven by ER (ESR1) gene amplification or overexpression