Characterization of the human blood coagulation factor X promoter.
Blood coagulation Factor X is a serine protease required for both the intrinsic and extrinsic pathways of coagulation. The gene for Factor X spans 27 kilobases and is located on chromosome 13, in close proximity to the gene encoding Factor VII. Expression of Factor X is restricted to the liver. We have characterized the human Factor X promoter by mapping the start sites of transcription and carrying out a functional analysis of the promoter. The first 279 base pairs (bp) of 5'-flanking sequence upstream from the first AUG are sufficient to confer maximal promoter activity in HepG2 cells. Protein-binding sites within the 279-bp fragment are defined using gel mobility shift assays. Mutagenesis of two specific sequences within the 279-bp fragment (CCAAT at -120 to -116, and ACTTTG at -56 to -51), results in loss of ability to bind proteins from a HepG2 nuclear extract, and profound reduction in promoter activity of the 279-bp fragment. We conclude that these two protein-binding sites are critical for the activity of the Factor X promoter.
Huang
M N
MN
Department of Medicine, School of Medicine, University of North Carolina, Chapel Hill 27599.
Hung
H L
HL
Stanfield-Oakley
S A
SA
High
K A
KA
D001483:Base Sequence; D001665:Binding Sites; D006528:Carcinoma, Hepatocellular; D002460:Cell Line; D002467:Cell Nucleus / Q000502:physiology; D002882:Chromosomes, Human, Pair 13; D005165:Factor V / Q000235:genetics*; D013006:Growth Hormone / Q000235:genetics; D006367:HeLa Cells; D006801:Humans; D008113:Liver Neoplasms; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D009838:Oligodeoxyribonucleotides; D010957:Plasmids; D016133:Polymerase Chain Reaction; D011401:Promoter Regions, Genetic; D015183:Restriction Mapping; D014162:Transfection
D009838:Oligodeoxyribonucleotides; D005165:Factor V; D013006:Growth Hormone