Alterations in DNA repair play major roles in breast cancer resistance to ionizing radiation (IR) and chemotherapies. Factors operating in both DSB repair and RNA transcription, specifically proteins regulating RNA polymerase (Pol) II, have been suggested, but few reported. When RNA Pol II stalls, or when transcription cannot properly terminate, RNA/DNA hybrids (R-loops) have extended half-lives that result in DNA double strand break (DSB) formation and genetic instability. Using yeast two hybrid analyses, we identified Ku70 binding protein #5 [(Kub5)/Hera], a highly conserved human homolog of yeast rtt103 that terminates RNA transcription via its regulation of RNA Pol II. Stable loss of Kub5/Hera expression, either by heterozygote knockout or knockdown by siRNA/shRNA expression, leads to elevated basal DSBs, ATM activation, foci formation and chromatid aberrations that were abrogated by RNase H forced over- expression, suggesting a role for R-loops in genetic instability. Kub5/Hera knockdown cells were hypersensitive to IR, equivalent to Ku70 deficient cells, and failed to repair DSBs that require DNA end-processing. We hypothesize that KUB5/HERA plays dual roles in the cell to: (i) regulate RNA Pol II to terminate RNA transcription via its interaction with the RNA Pol II CTD domain;and (ii) stimulate NHEJ processing of complex DSBs by Ku70 and Artemis interactions. When its expression is depressed (e.g., one-half via haplo-insufficiency) R-loops form and DSBs are created due to deficient RNA transcription termination. DNA damage is simultaneously amplified by the cell's inability to repair DSBs downstream, leading to IR hypersensitivity.
Three Specific Aims are proposed:
Aim 1 : to perform structure/function analyses of Kub5/Hera focusing on two regions, the RPR and coiled-coil domains, to uncouple transcription termination and DSB repair;
Aim 2 : to define functions of KUB5- Ku70-Artemis complexes in DSB repair;
and Aim 3 : to define the role(s) of KUB5/HERA in regulating RNA Pol II function, RNA transcription termination, and spontaneous DSB formation and chromatid aberrations. These studies will allow us to explore the role of KUB5/HERA over-expression in human breast cancer radio-resistance.

Public Health Relevance

Human Ku70 binding protein #5, also known as HERA, (KUB5/HERA) is a new factor involved in the repair of DNA ends with complex damage, such as blunt-end breaks. Elevations of KUB5/HERA in breast and ovarian cancers, and our demonstration that KUB5/HERA is extremely important for drug and radiation resistance, suggests that this protein is a valuable target for future therapies and a 'valuable predictor of response'for therapies. Our recent observation that KUB5/HERA is required for RNA termination allows us to also investigate the interface between DNA repair and regulation of RNA Polymerase II and transcription termination.

National Institute of Health (NIH)
National Cancer Institute (NCI)
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Special Emphasis Panel (ZRG1-OTC-K (06))
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Ogunbiyi, Peter
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University of Texas Sw Medical Center Dallas
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Jiang, L; Xiao, L; Sugiura, H et al. (2015) Metabolic reprogramming during TGF?1-induced epithelial-to-mesenchymal transition. Oncogene 34:3908-16
Morales, Julio; Li, Longshan; Fattah, Farjana J et al. (2014) Review of poly (ADP-ribose) polymerase (PARP) mechanisms of action and rationale for targeting in cancer and other diseases. Crit Rev Eukaryot Gene Expr 24:15-28
Fattah, Farjana J; Hara, Kodai; Fattah, Kazi R et al. (2014) The transcription factor TFII-I promotes DNA translesion synthesis and genomic stability. PLoS Genet 10:e1004419
Luo, Xiuquan; Suzuki, Masatoshi; Ghandhi, Shanaz A et al. (2014) ATM regulates insulin-like growth factor 1-secretory clusterin (IGF-1-sCLU) expression that protects cells against senescence. PLoS One 9:e99983
Li, L; Chakraborty, S; Yang, C-R et al. (2014) An EGFR wild type-EGFRvIII-HB-EGF feed-forward loop regulates the activation of EGFRvIII. Oncogene 33:4253-64
Morales, Julio C; Richard, Patricia; Rommel, Amy et al. (2014) Kub5-Hera, the human Rtt103 homolog, plays dual functional roles in transcription termination and DNA repair. Nucleic Acids Res 42:4996-5006
Chakraborty, Sharmistha; Li, Li; Tang, Hao et al. (2013) Cytoplasmic TRADD confers a worse prognosis in glioblastoma. Neoplasia 15:888-97
Puliyappadamba, Vineshkumar Thidil; Chakraborty, Sharmistha; Chauncey, Sandili S et al. (2013) Opposing effect of EGFRWT on EGFRvIII-mediated NF-*B activation with RIP1 as a cell death switch. Cell Rep 4:764-75
Klokov, D; Leskov, K; Araki, S et al. (2013) Low dose IR-induced IGF-1-sCLU expression: a p53-repressed expression cascade that interferes with TGFýý1 signaling to confer a pro-survival bystander effect. Oncogene 32:479-90