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Author Contact: ekim03@wesleyan.edu
Local Identifier: 1701"},{"label":"Author-supplied Keywords","value":["Metal-binding site","Single-nucleotide polymorphism","Cervical cancer cells","Zinc transport","ZIP11"]},{"label":"Degree","value":["Master of Arts"]},{"label":"Description","value":"

Zinc (Zn) is one of the most abundant elements in the environment, and an essential micronutrient that regulates catalytic function of enzymes, gene expression, signaling and confers structural stability to proteins. Zn exists in an inactive redox state as a divalent cation (Zn2+) under physiological conditions, making it key in biological processes. Maintaining Zn homeostasis is crucial in controlling normal cell development and function, thus dysregulation of supply and transport leads to pathophysiological conditions. Zinc transporters are essential regulators of Zn homeostasis. Among these, ZIP11 is a member of the gufA subfamily of ZIP transporters in mammals and is localized to the nucleus and the Golgi apparatus. Our lab discovered that ZIP11 maintains nuclear Zn levels in mammalian cells. To investigate the role of ZIP11 in cervical cancer, we stably knocked down this transporter in HeLa cells and found that this led to nuclear Zn accumulation, reducing cell proliferation and impairing cell migration and invasive properties. Reintroducing ZIP11 rescued the phenotype, while overexpression in wild type HeLa cells led to a more aggressive carcinogenic phenotype. Genomic analysis of publicly available cervical and ovarian cancer patients’ datasets identified several single-nucleotide polymorphisms (SNPs) in the ZIP11 coding region that may represent potential deleterious consequences. Due to the location of these SNPs, we hypothesized that these mutations may affect ZIP11 function by increasing substrate accessibility, potentially enhancing the carcinogenic phenotype of HeLa cells. On the other hand, we also identified some key residues involved in transmembrane metal transport, and further hypothesized that mutations of these metal-binding sites (MBS) may impair Zn mobilization, rendering a less carcinogenic phenotype. The purpose of my research was to evaluate the biological effects of introducing single SNPs and MBS mutations to the ZIP11 transporter. KD ZIP11 HeLa cells reconstituted with ZIP11 encoding into the single SNPs showed restored carcinogenic properties and mitochondrial potential, while cells expressing ZIP11 with the MBS mutations presented a similar phenotype to KD cells. Understanding the effects of SNPs and MBS mutations on the function of ZIP11 in maintaining nuclear zinc homeostasis in cervical cancer cells will help to elucidate the role of the transporter in cancer progression.

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