Site-directed mutagenesis reveals a liver transcription factor essential for the albumin transcriptional enhancer.
A method based on the polymerase chain reaction is described for constructing a clustered set of basepair changes, deletions, or insertions at any site on a DNA fragment. Advantages of the procedure are that virtually every product has the desired sequence alteration and that only a single round of polymerase chain reaction is required. We used this method to demonstrate that the binding of a specific liver nuclear protein, which we call eH-TF, is essential for the function of the enhancer of the mouse albumin gene. The eH-TF binding activity is hepatocyte-specific; it binds to a functional region of the albumin promoter and is distinct from other albumin promoter factors, and part of the eH-TF binding sequence, TGTTTGC, occurs in functional regulatory sites of other liver-specific genes.
Zaret
K S
KS
Section of Biochemistry, Brown University, Providence, RI 02912.
Liu
J K
JK
DiPersio
C M
CM
D000818:Animals; D001483:Base Sequence; D002467:Cell Nucleus / Q000378:metabolism; D003850:Deoxyribonuclease I; D004742:Enhancer Elements, Genetic; D008099:Liver / Q000378:metabolism*; D051379:Mice; D008969:Molecular Sequence Data; D009154:Mutation; D009687:Nuclear Proteins / Q000235:genetics*; D015345:Oligonucleotide Probes; D016133:Polymerase Chain Reaction; D011401:Promoter Regions, Genetic; D015183:Restriction Mapping; D012709:Serum Albumin / Q000235:genetics*; D014157:Transcription Factors / Q000235:genetics*; D014158:Transcription, Genetic
D009687:Nuclear Proteins; D015345:Oligonucleotide Probes; D012709:Serum Albumin; D014157:Transcription Factors; D003850:Deoxyribonuclease I