The tumor suppressor p53 is a critical sensor of cellular DNA damage. Cellular stress is signaled to p53 by a number of different signal transduction pathways. We recently identified that DMC activates both p53 dependent and independent apoptotic pathways while MC only activates the p53 dependent pathway. Understanding how chemotherapeutic drugs signal to wild-type p53 is of central relevance to cancer treatment. Determining DNA damage pathways that can signal to p53 family members, p73 and p63 has emerged as equally important with the recent finding that these proteins are required to coordinately regulate the DNA damage apoptotic response. Our hypothesis is that DMC can significantly activate a p53-independent apoptotic pathway that utilizes p73 and p63. The long-range goal is to determine the p53-independent signal transduction pathway(s) activated by DMC.
Aim 1) Identify the cytotoxicity and specific target gene regulation, and checkpoint kinases activated, by DMC as compared to other DNA damaging drugs in cell lines, with and without p53. The cytotoxicity and the expression of selected target genes known to associate with the induction of apoptosis and cell cycle regulation will be determined by drug dose curves of representative cell lines with MC, DMC and a battery of well characterized chemotherapeutics. Gene expression levels of multiple validated targets (including Bax, Fas, Puma, Noxa, Gadd45, p21 and mdm2) will be monitored using real-time RT-PCR and target kinase activation will be monitored by phosphor-specific antibodies.
Aim 2) Determine if differential activation of p53 target genes associates with alternative multiprotein complex formation on chromatin. Chromatin immuno-precipitation-PCR (ChlP-PCR), utilizing p53 specific, p73 specific and MDM2 specific antibodies, will be used to investigate the nuclear in vivo interaction of p53, p73 and MDM2 at the known enhancer binding sites for the apoptotic and cell cycle regulating target genes. Fixation is the first step in the ChlP-PCR protocol and will preserve higher-molecular weight complexes.
Aim 3) Examine gene expression by affymetrix DNA micro-arrays in MC and DMC treated cell lines with and without p53 as well as with and without the p53 dependent apoptosis inhibitor WISP-1. To determine whether apoptosis can be attributed to specific overexpressed genes, siRNA will be used for functional analysis.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Minority Biomedical Research Support - MBRS (S06)
Project #
5S06GM060654-08
Application #
7391697
Study Section
Minority Programs Review Committee (MPRC)
Project Start
Project End
Budget Start
2007-04-01
Budget End
2008-03-31
Support Year
8
Fiscal Year
2007
Total Cost
$220,035
Indirect Cost
Name
Hunter College
Department
Type
DUNS #
620127915
City
New York
State
NY
Country
United States
Zip Code
10065
Luine, Victoria; Gomez, Juan; Beck, Kevin et al. (2017) Sex differences in chronic stress effects on cognition in rodents. Pharmacol Biochem Behav 152:13-19
Gupta, Rupal; Huang, Wenlin; Francesconi, Lynn C et al. (2017) Effect of positional isomerism and vanadium substitution on 51V magic angle spinning NMR Spectra Of Wells-Dawson polyoxotungstates. Solid State Nucl Magn Reson 84:28-33
Luine, Victoria (2016) Estradiol: Mediator of memories, spine density and cognitive resilience to stress in female rodents. J Steroid Biochem Mol Biol 160:189-95
Luine, Victoria (2015) Recognition memory tasks in neuroendocrine research. Behav Brain Res 285:158-64
Frankfurt, Maya; Luine, Victoria (2015) The evolving role of dendritic spines and memory: Interaction(s) with estradiol. Horm Behav 74:28-36
DeCicco, Jennifer M; O'Toole, Laura J; Dennis, Tracy A (2014) The late positive potential as a neural signature for cognitive reappraisal in children. Dev Neuropsychol 39:497-515
Luine, Victoria N (2014) Estradiol and cognitive function: past, present and future. Horm Behav 66:602-18
Garcia, Miguel; Ray, Sibnath; Brown, Isaiah et al. (2014) PakD, a putative p21-activated protein kinase in Dictyostelium discoideum, regulates actin. Eukaryot Cell 13:119-26
O'Toole, Laura J; DeCicco, Jennifer M; Berthod, Samantha et al. (2013) The N170 to angry faces predicts anxiety in typically developing children over a two-year period. Dev Neuropsychol 38:352-63
Garcia, Rebecca; Nguyen, Liem; Brazill, Derrick (2013) Dictyostelium discoideum SecG interprets cAMP-mediated chemotactic signals to influence actin organization. Cytoskeleton (Hoboken) 70:269-80

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