Open AccessDeterminants of the brain‐specific expression of the rat aldolase C gene: ex vivo and in vivo analysis
Author(s) -
THOMAS Muriel,
MAKEH Iman,
BRIAND Pascale,
KAHN Axel,
SKALA Henriette
Publication year - 1993
Publication title -
european journal of biochemistry
Language(s) - English
Resource type - Journals
eISSN - 1432-1033
pISSN - 0014-2956
DOI - 10.1111/j.1432-1033.1993.tb18360.x
Subject(s) - biology , microbiology and biotechnology , gene , caat box , promoter , mutation , transgene , reporter gene , aldolase a , genetics , ex vivo , gene expression , in vivo , enzyme , biochemistry
A 115‐bp promoter fragment of the aldolase C gene is sufficient for conferring neural cell specificity on a reporter gene, in cultured PC12 cells and in transgenic mice. In vitro DNase I protection experiments detected two footprints on the promoter, termed boxes A/A', and B. The 5′ A/A' box contains overlapping Sp1 and Krox20/Krox24 binding sites; it binds Sp1 in fibroblasts (box A') and a different complex in brain (box A). Any deletion or mutation of this box that impairs protein recognition also suppresses promoter activity. The replacement of box A/A' by a Sp1 consensus binding site results in the loss of the brain specificity of expression in transgenic mice. Further 3′, box B is composed of a 5′ direct repeat and a 3′ GC box consisting of overlapping Sp1 and Krox20/Krox24 binding sites. Mutation of the direct repeat subregion appears to be more deleterious for the promoter activity than mutation of the G+C‐rich subregion.