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Page 1: P53- and Drug-Induced Apoptotic Responses Mediated by BH3-Only Proteins Puma and Noxa P53- and Drug-Induced Apoptotic Responses Mediated by BH3-Only Proteins

P53- and Drug-Induced P53- and Drug-Induced Apoptotic Responses Mediated Apoptotic Responses Mediated by BH3-Only Proteins Puma by BH3-Only Proteins Puma

and Noxaand Noxa

Villunger, A. et al. Science, 2003

Flávia Gama Gomes Leite

Page 2: P53- and Drug-Induced Apoptotic Responses Mediated by BH3-Only Proteins Puma and Noxa P53- and Drug-Induced Apoptotic Responses Mediated by BH3-Only Proteins

p53 – ciclo celular e p53 – ciclo celular e apoptoseapoptose

p53

p21

Parada do ciclo celular

Mais de 16 genes alvos

Apoptose ?

Page 3: P53- and Drug-Induced Apoptotic Responses Mediated by BH3-Only Proteins Puma and Noxa P53- and Drug-Induced Apoptotic Responses Mediated by BH3-Only Proteins

Possíveis Genes AlvosPossíveis Genes Alvos

Puma (p53-upregulated modulator of apoptosis)

Noxa (Noxius stress)

Domínio BH3

Page 4: P53- and Drug-Induced Apoptotic Responses Mediated by BH3-Only Proteins Puma and Noxa P53- and Drug-Induced Apoptotic Responses Mediated by BH3-Only Proteins

Construção de NoxaConstrução de Noxa-/--/-

Page 5: P53- and Drug-Induced Apoptotic Responses Mediated by BH3-Only Proteins Puma and Noxa P53- and Drug-Induced Apoptotic Responses Mediated by BH3-Only Proteins

NoxaNoxa-/--/-

Northern blot

Agarose - controle

Southern blot

PCR

Southern blot

Page 6: P53- and Drug-Induced Apoptotic Responses Mediated by BH3-Only Proteins Puma and Noxa P53- and Drug-Induced Apoptotic Responses Mediated by BH3-Only Proteins

NoxaNoxa-/--/-

Estímulo dependente de p53

Page 7: P53- and Drug-Induced Apoptotic Responses Mediated by BH3-Only Proteins Puma and Noxa P53- and Drug-Induced Apoptotic Responses Mediated by BH3-Only Proteins

NoxaNoxa-/--/-

Estímulos independentes de p53

Estímulos dependentes de p53

Page 8: P53- and Drug-Induced Apoptotic Responses Mediated by BH3-Only Proteins Puma and Noxa P53- and Drug-Induced Apoptotic Responses Mediated by BH3-Only Proteins

Construção de PumaConstrução de Puma-/--/-

Page 9: P53- and Drug-Induced Apoptotic Responses Mediated by BH3-Only Proteins Puma and Noxa P53- and Drug-Induced Apoptotic Responses Mediated by BH3-Only Proteins

PumaPuma-/--/-Southern blot

PCR

Southern blot - sonda exon 2

RT- PCR

Northen blot

Page 10: P53- and Drug-Induced Apoptotic Responses Mediated by BH3-Only Proteins Puma and Noxa P53- and Drug-Induced Apoptotic Responses Mediated by BH3-Only Proteins

PumaPuma-/--/-

genotóxico

Page 11: P53- and Drug-Induced Apoptotic Responses Mediated by BH3-Only Proteins Puma and Noxa P53- and Drug-Induced Apoptotic Responses Mediated by BH3-Only Proteins

Independente de p53

Page 12: P53- and Drug-Induced Apoptotic Responses Mediated by BH3-Only Proteins Puma and Noxa P53- and Drug-Induced Apoptotic Responses Mediated by BH3-Only Proteins

PumaPuma-/--/-

Page 13: P53- and Drug-Induced Apoptotic Responses Mediated by BH3-Only Proteins Puma and Noxa P53- and Drug-Induced Apoptotic Responses Mediated by BH3-Only Proteins

Puma-/-


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