Direct α-synuclein promoter transactivation by the tumor suppressor p53

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Transcript of Direct α-synuclein promoter transactivation by the tumor suppressor p53

  • Tema 1

    PRESENTED BY:

    Ana Carolina Betancur Echavarra Mabel Dahiana Roldan Tabares

    2016

  • Cancer: Cancer is defined like abnormal

    cells divide in an uncontrolled way.

    It starts when a gen changes,

    make one cell or a few cells begin to grow and divide too much. This may cause a growth called a tumor.

    Cancer may be associated with

    deficit of p53

  • p53: Its a protein that generates an

    answer when the cell suffers an injury.

    p53 regulates the interphase through a checkpoint between G1/S phases.

    This protein can cause:

    oDNA reparation.

    o Apoptosis.

    o Cell cycle arrest.

  • Parkinson disease: Its a motor disease associated with the

    degeneration of dopaminergic neurons of the substantia nigra pars compacta.

    With less dopamine, a person has less and less

    ability to regulate their movements, body and emotions.

    In which several causative genes have been

    identified. Amongst -synuclein, DJ-1 and parkin.

    In most cases PD is associated with Lewy bodies that is an accumulation of -Synuclein.

  • -Synuclein: Presinpatic neuronal protein involved

    both genetically and neuropathologically with Parkinsons disease.

    Its abundantly expressed in the nervous system.

    Levels of -Synuclein are modulated when there are conditions that alter plasticity or confer injury.

    It has the ability to down-regulate p53.

  • From: -Synuclein and Parkinsons disease. Disponible at:http://www.fasebj.org/content/18/6/617/F2.large.jpg

  • Dogma and relation:

  • To show that p53 could also control -syn levels as part of a feedback process driving their cellular homeostasis in neurons.

    To show that the disruption of cellular dialogue between p53 and -syn linked to the dysfunction of any of these two partners may contribute to PD pathology.

  • CLULAS

    Fibroblastos embrionarios de ratn.

    Adenocarcinoma colorrectal humano.

    SH-SY5Y (lnea celular de neuroblastoma humano).

    Clulas HAP1 (lnea celular haploide humana).

    Modelos animales y celulares.

  • Modulacin del p53 a travs de frmacos y radiaciones UV.

    Leptomicina provoca la detencin del ciclo celular G1 en clulas de mamferos, adems de inhibir la

    exportacin nuclear.

    El etopsido

  • Western blot: Para identificar sinuclena, p53 y actina.

  • Transfeccin a travs de plsmidos.

  • Ensayo basado en el reporte de Luciferasa.

    Identificacin del gen de inters Clonacin. Gen de inters + luciferasa agregados a un plsmido Transfeccin La luciferasa se transcribir proporcionalmente a cmo lo

    hara el gen de inters. Cultivo las clulas y las liso, dejando la luciferasa libre.

  • PCR Amplificar

    genes -syn y su promotor.

  • EMSA. Para evaluar la interaccin entre el gen que codifica para -

    syn y p53 en ausencia de cualquier modulador o intermediario.

  • Ensayo de inmunoprecipitacin de cromatina

  • Transfeccin vector vacio

    Transfeccin vector con DNAc de p53

  • EV: Vector vaco P53: Vectores que codifican para P53

  • AUTHOR WHAT DID HE SAY? AGREE DISAGREE

    Vousden KH. p53 is a trascriptional factor , which main physiological function is to regulated genes involved in the

    control of cell cycle, DNA repair and apoptosis.

    Checler F. p53 is a proapoptotic protein strongly linked to several neurodegenerative

    disorders, amongst which Parkinsons disease.

    Checler F. PD-causative gene products including DJ-1 and parkin, have been shown to control and to be controlled by p53

    ex-vivo and in vivo.

    Alves da Costa C.

    In normal conditions, -syn represses p53.

  • The identification of Synuclein as a main target protein regulated by p53 may offer a new perspective about new treatments for PD that leads to delay the progress of this disease.

    Knowledge about the interaction between Synuclein and p53 allows to further new studies in order to identify eficient ways to generate either up or down regulation of those proteins.

  • Knowing the behavior of a-syn and p53 on feedback allows us focusing the treatment scheme of Parkinson disease to be more effective and with less contraindications.

    The development of Parkinson's disease, its given be multiple causes, for example the use of certain drugs, exposure to external agents. You might even be associated with other diseases such as cancer. So it is only the first step to diagnose new causes.

  • Stefanis L. -Synuclein in Parkinsons Disease. CSH Perspectives.

    2012 (citado 28 de febrero, 2016). Cold Spring Harb Perspect Med 2012;4:a009399. Disponible en: http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3281589/pdf/cshperspectmed-PKD-a009399.pdf

    Weber JD, Jeffers JR, Rehg EJ, Randle DH, Lozano G, Roussel MF et al. p53-independent functions of the p19ARF tumor suppressor. CSH Perspectives [Internet]. 2000 (citado 28 de febrero, 2016). Genes & Dev. 2000. 14: 2358-2365. Disponible en: http://genesdev.cshlp.org/content/14/18/2358.full.pdf+html

  • Castro Toro A, Buritic OF. Enfermedad de parkinson: criterios

    diagnsticos, factores de riesgo y de progresin, y escalas de valoracin del estadio clnico. Acta Neurol Colomb Internet]. 2014 (citado 01 de marzo,2016). Vol. 30(4):300-306. Disponible en: http://www.scielo.org.co/pdf/anco/v30n4/v30n4a10.pdf

    Alves da Costa C, Paitel E, Vincent B, Checler F. JBC [Internet]. -Synuclein Lowers p53-dependent Apoptotic Response of Neuronal Cells. 2002 (citado 01 de marzo,2016). Vol. 277, No. 52, pp. 5098050984. Disponible en: http://www.jbc.org/content/277/52/50980.full.pdf+html

  • CMAP: Ana Carolina.

  • CMAP: MABEL