Figure 1.
Identification of essentialcis-actingelements within thec1orf109promoter.(A) DNA sequence analysis of the 5′ flanking region of human c1orf109 gene. The nucleotide sequence is numbered with the transcriptional start site (TSS) as +1 (dark arrow). MatInspector online software was used to predict the putative transcription factor binding sites, which are underlined and indicated by name. (B) Deletion analysis of the c1orf109 promoter. The truncated promoter fragments were inserted into the luciferase reporter vector pGL3-basic (Luc). Approximately 2 × 105 HEK293 in each well of 24-well plate were transfected with 1.0 μg each of pGL3-c1orf109 promoter constructs plus 50 ng of phRL-SV40 vector. The firefly luciferase activity was assayed 24 hr after transfection and normalized to Renilla luciferase activity. Values are represented by means ± S.D. obtained from three independent experiments. Deletion from −177 to −101 bp and −101 to −41 bp led to extreme reduction of activity. The region within −177 bp of the c1orf109 promoter contains multiple potentially important transcription factor binding sites including CAAT and TATA boxes.