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

p53 – ciclo celular e p53 – ciclo celular e apoptoseapoptose

p53

p21

Parada do ciclo celular

Mais de 16 genes alvos

Apoptose ?

Possíveis Genes AlvosPossíveis Genes Alvos

Puma (p53-upregulated modulator of apoptosis)

Noxa (Noxius stress)

Domínio BH3

Construção de NoxaConstrução de Noxa-/--/-

NoxaNoxa-/--/-

Northern blot

Agarose - controle

Southern blot

PCR

Southern blot

NoxaNoxa-/--/-

Estímulo dependente de p53

NoxaNoxa-/--/-

Estímulos independentes de p53

Estímulos dependentes de p53

Construção de PumaConstrução de Puma-/--/-

PumaPuma-/--/-Southern blot

PCR

Southern blot - sonda exon 2

RT- PCR

Northen blot

PumaPuma-/--/-

genotóxico

Independente de p53

PumaPuma-/--/-

Puma-/-