Open AccessDNA bending and binding factors of the human β-actin promoter
Author(s) -
Takeshi Kawamoto,
Kozo Makino,
Satoshi Orita,
Atsuo Nakata,
Takeo Kakunaga
Publication year - 1989
Publication title -
nucleic acids research
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 9.008
H-Index - 537
eISSN - 1362-4954
pISSN - 0305-1048
DOI - 10.1093/nar/17.2.523
Subject(s) - biology , microbiology and biotechnology , homology (biology) , dna , transcription (linguistics) , chloramphenicol acetyltransferase , promoter , gene , deoxyribonuclease i , conserved sequence , actin , regulatory sequence , transcription factor , gene expression , genetics , peptide sequence , base sequence , philosophy , linguistics
Transcription of the beta-actin gene is rapidly inducible in response to serum stimulation. To determine the regions responsible for serum inducible and basal level expression, the human beta-actin promoter was subjected to mutational analysis. Two distinct elements, the CCAAT homology and the beta-actin specific conserved sequences, were found by a chloramphenicol acetyltransferase expression assay and sequence comparisons, and then analyzed for possible functions. Using a DNA bend assay, it was shown that the conserved sequences included the core of a sequence-directed bend of DNA. Gel mobility shift and DNase I protection assays revealed that the conserved sequences and the CCAAT homology were recognized by binding factors in HeLa cell extracts.