Articles by Joel F Andrews in JoVE
Application of Laser Micro-irradiation for Examination of Single and Double Strand Break Repair in Mammalian Cells Nathaniel W Holton1, Joel F Andrews1, Natalie R Gassman1 1Department of Oncologic Sciences, University of South Alabama Mitchell Cancer Institute Confocal fluorescence microscopy and laser micro-irradiation offer tools for inducing DNA damage and monitoring the response of DNA repair proteins in selected sub-nuclear areas. This technique has significantly advanced our knowledge of damage detection, signaling, and recruitment. This manuscript demonstrates these technologies to examine single and double strand break repair.
Other articles by Joel F Andrews on PubMed
Cellular Stress Stimulates Nuclear Localization Signal (NLS) Independent Nuclear Transport of MRJ Experimental Cell Research. Jun, 2012 | Pubmed ID: 22504047 HSP40 family member MRJ (DNAJB6) has been in the spot light for its relevance to Huntington's, Parkinson's diseases, limb-girdle muscular dystrophy, placental development, neural stem cells, cell cycle and malignancies such as breast cancer and melanoma. This gene has two spliced variants coding for 2 distinct proteins with significant homology. However, MRJ(L) (large variant) is predominantly localized to the nucleus whereas MRJ(S) (small variant) is predominantly cytoplasmic. Interestingly MRJ(S) translocates to the nucleus in response to heat shock. The classical heat shock proteins respond to crises (stress) by increasing the number of molecules, usually by transcriptional up-regulation. Our studies imply that a quick increase in the molar concentration of MRJ in the nuclear compartment is a novel method by which MRJ responds to stress. We found that MRJ(S) shows NLS (nuclear localization signal) independent nuclear localization in response to heat shock and hypoxia. The specificity of this response is realized due to lack of such response by MRJ(S) when challenged by other stressors, such as some cytokines or UV light. Deletion analysis has allowed us to narrow down on a 20 amino acid stretch at the C-terminal region of MRJ(S) as a potential stress sensing region. Functional studies indicated that constitutive nuclear localization of MRJ(S) promoted attributes of malignancy such as proliferation and invasiveness overall indicating distinct phenotypic characteristics of nuclear MRJ(S).
Suppression of Ser/thr Phosphatase 4 (PP4C/PPP4C) Mimics a Novel Post-mitotic Action of Fostriecin, Producing Mitotic Slippage Followed by Tetraploid Cell Death Molecular Cancer Research : MCR. May, 2013 | Pubmed ID: 23671329 Fostriecin is a natural product purified from Sterptomyces extracts with antitumor activity sufficient to warrant human clinical trials. Unfortunately, difficulties associated with supply and stable drug formulation stalled further development. At a molecular level, fostriecin is known to act as a catalytic inhibitor of four PPP-family-phosphatases, and reports describing the syntheses of designed molecules in the class suggest derivatives targeting enzymes within the fostriecin sensitive sub-family can be produced. However, it is not clear if the tumor selective cytotoxicity of fostriecin results from the inhibition of a specific phosphatase, multiple phosphatases, or a limited subset of fostriecin sensitive phosphatases. How the inhibition of sensitive phosphatases contributes to tumor selective cytotoxicity is also not clear. Here, high-content time-lapse imaging of live cells reveals novel insight into the cellular actions of fostriecin, showing that fostriecin induced apoptosis is not simply induced following a sustained mitotic arrest. Rather, apoptosis occurs in an apparent second interphase produced when tetraploid cells undergo mitotic slippage. Comparison of the actions of fostriecin and antisense-oligonucleotides specifically targeting human fostriecin-sensitive phosphatases revealed that the suppression PP4C alone is sufficient to mimic many actions of fostriecin. Importantly, antisense-oligonucleotides targeting PP4C induce apoptosis, with death occurring in tetraploid cells produced following mitotic slippage. This affect was not observed following the suppression of PP1C, PP2AC or PP5C. Although future studies are needed to clarify how the suppression of PP4C triggers mitotic slippage/apoptosis, our observations suggest further development of fostriecin class inhibitors should consider PP4C as a potentially important target.