Project 1 will focus on the pathways of DMA repair that are known to participate in cellular responses to UVinduced DMA damage: nucleotide excision repair (NER), post-replication repair (PRR) and DMA doublestrand break (dsb) repair.
Specific aim 1 will investigate functional NER capacity in melandcytes derived from healthy Caucasian newborns, in melanoma cell lines, and in melanocytes with genetic alterations commonly found in melanoma. The main goal is to determine whether NER capacity is attenuated or lost during stages of development of melanoma. In addition, aim 1 will include in vitro assays with purified proteins for a mechanistic test of the kinetic proofreading model of NER.
Specific Aim 2 will determine the capacity of melanocytes and melanoma lines to tolerate unrepaired DNA phptoproducts during replication. It will examine an innovative new hypothesis that UV-induced DNA damage triggers a signaling pathway that results in trans-inhibition of DNA chain elongation in active replicons. This is achieved by the active reduction in the rate of progression of DNA replication forks before the direct encounter with a template lesion. A DNA fiber-combing and immuno-staining assay will enable visualization of replication dynamics in individual replicons. Knockdown of Timeless and Tipin, among other proteins of interest, will determine whether the replication fork protection complex regulates the rate of displacement of DNA replication forks in UV-damaged cells.
Specific aim 3 will examine the induction of chromosomal aberrations and allelic deletions in UV-treated cells. Chromosomal aberrations are thought to be associated with DNA dsb generated at collapsed replication forks and other single-strand DNA regions formed during replication of the UV-damaged DNA. Phospho-histone H2AX/phosphd-ATM/MRE11 -positive nuclear foci will be quantified to monitor the formation and repair of DNA dsb. Studies will determine whether genetic alterations that induce melanoma in reconstructed human skin enhance UV-clastogenesis in cultured melanocytes. This project wil determine whether defects in DNA repair produce a UV-chromosomal-mutator phenotype in skin melanocytes.
| Cordeiro-Stone, Marila; McNulty, John J; Sproul, Christopher D et al. (2016) Effective intra-S checkpoint responses to UVC in primary human melanocytes and melanoma cell lines. Pigment Cell Melanoma Res 29:68-80 |
| Lin, Ja-An; Zhu, Hongtu; Mihye, Ahn et al. (2014) Functional-mixed effects models for candidate genetic mapping in imaging genetic studies. Genet Epidemiol 38:680-91 |
| Nikolaishvilli-Feinberg, Nana; Cohen, Stephanie M; Midkiff, Bentley et al. (2014) Development of DNA damage response signaling biomarkers using automated, quantitative image analysis. J Histochem Cytochem 62:185-96 |
| Kaufmann, William K; Carson, Craig C; Omolo, Bernard et al. (2014) Mechanisms of chromosomal instability in melanoma. Environ Mol Mutagen 55:457-71 |
| Chen, Liddy M; Ibrahim, Joseph G; Chu, Haitao (2014) Flexible stopping boundaries when changing primary endpoints after unblinded interim analyses. J Biopharm Stat 24:817-33 |
| Kricker, Anne; Armstrong, Bruce K; Goumas, Chris et al. (2013) Survival for patients with single and multiple primary melanomas: the genes, environment, and melanoma study. JAMA Dermatol 149:921-7 |
| Schlegel, Jennifer; Sambade, Maria J; Sather, Susan et al. (2013) MERTK receptor tyrosine kinase is a therapeutic target in melanoma. J Clin Invest 123:2257-67 |
| Omolo, Bernard; Carson, Craig; Chu, Haitao et al. (2013) A prognostic signature of G(2) checkpoint function in melanoma cell lines. Cell Cycle 12:1071-82 |
| Smith-Roe, Stephanie L; Patel, Shivani S; Zhou, Yingchun et al. (2013) Separation of intra-S checkpoint protein contributions to DNA replication fork protection and genomic stability in normal human fibroblasts. Cell Cycle 12:332-45 |
| Lakhter, Alexander J; Sahu, Ravi P; Sun, Yang et al. (2013) Chloroquine promotes apoptosis in melanoma cells by inhibiting BH3 domain-mediated PUMA degradation. J Invest Dermatol 133:2247-54 |
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