ISOLATION OF THE MURINE BRANCHED-CHAIN ALPHA-KETOACID DEHYDROGENASE E2 SUBUNIT PROMOTER REGION

The promoter region of the murine branched-chain a-ketoacid dehydrogen ase E2 subunit (dihydrolipoyl transacylase) gene was isolated and char acterized. Sequence analysis of the promoter-regulatory region showed the presence of two inverted 'CAAT box' sequences, the most proximal b eing -42 to -48 bp upstream from the determined transcription initiati on site, but no TATA-box sequences, similar to the human BCKAD E2 gene . The boundary of the minimum promoter sequence appeared to reside jus t inclusive of this first inverted CAAT sequence, but minigene transfe r analysis demonstrated that the promoter proximal region between -65 and -140 bp is likely to be extremely important for controlling regula ted changes in E2 RNA expression, The regulatory effect of this region may be modulated by a number of other upstream regions which were ide ntified within the 7.0 kb sequence examined. (C) 1998 Elsevier Science B.V. All rights reserved.