Candi E, Rufini A, Terrinoni A, Dinsdale D, Ranalli M, Paradisi A, De Laurenzi V, Spagnoli L G, Catani M V, Ramadan S, Knight R A, Melino G
Biochemistry Laboratory, IDI-IRCCS, c/o University of Rome Tor Vergata, 00133 Rome, Italy.
Cell Death Differ. 2006 Jun;13(6):1037-47. doi: 10.1038/sj.cdd.4401926.
Epidermal development requires the transcription factor p63, as p63-/- mice are born dead, without skin. The gene expresses two proteins, one with an amino-terminal transactivation domain (TAp63) and one without (deltaNp63), although their relative contribution to epidermal development is unknown. To address this issue, we reintroduced TAp63alpha and/or deltaNp63alpha under the K5 promoter into p63-/- mice by in vivo genetic complementation. Whereas p63-/- and p63-/-;TA mice showed extremely rare patches of poorly differentiated keratinocytes, p63-/-;deltaN mice showed significant epidermal basal layer formation. Double TAp63alpha/deltaNp63alpha complementation showed greater patches of differentiated skin; at the ultrastructural level, there was clear reformation of a distinct basal membrane and hemidesmosomes. At the molecular level, deltaNp63 regulated expression of genes characteristic of the basal layer (K14), interacting (by Chip, luc assay) with the third p53 consensus site. Conversely, TAp63 transcribed the upper layer's genes (Ets-1, K1, transglutaminases, involucrin). Therefore, the two p63 isoforms appear to play distinct cooperative roles in epidermal formation.
表皮发育需要转录因子p63,因为p63基因敲除的小鼠出生时即死亡,且没有皮肤。该基因表达两种蛋白质,一种带有氨基末端反式激活结构域(TAp63),另一种则没有(deltaNp63),尽管它们对表皮发育的相对贡献尚不清楚。为了解决这个问题,我们通过体内基因互补技术,在K5启动子的作用下,将TAp63alpha和/或deltaNp63alpha重新导入p63基因敲除的小鼠体内。p63基因敲除小鼠和p63基因敲除;TA小鼠仅显示出极罕见的低分化角质形成细胞斑块,而p63基因敲除;deltaN小鼠则显示出明显的表皮基底层形成。TAp63alpha/deltaNp63alpha双互补显示出更大面积的分化皮肤斑块;在超微结构水平上,明显重新形成了独特的基底膜和半桥粒。在分子水平上,deltaNp63调节基底层特征基因(K14)的表达,并(通过芯片、荧光素酶检测)与第三个p53共有位点相互作用。相反,TAp63转录上层基因(Ets-1、K1、转谷氨酰胺酶、内披蛋白)。因此,这两种p63异构体在表皮形成过程中似乎发挥着不同的协同作用。