Characterization of a helicase-like transcription factor involved in the expression of the human plasminogen activator inhibitor-1 gene.
A 5.4-kb cDNA encoding the protein that binds to the B Box of the plasminogen activator inhibitor-1 (PAI-1) gene was isolated and sequenced. The protein, named helicase-like transcription factor (HLTF), contains a DNA-binding domain, a RING finger domain, and seven helicase domains and is homologous to SWI/SNF proteins. Two HLTF mRNAs of 5.5 and 4.5 kb were detected in most human tissues, a single gene was located on chromosome 3q24-25, and the protein was located in the nucleoplasm. Two HLTF proteins differing in translation start site (Met-1 or Met-123) were obtained by in vitro translation in reticulocyte lysate or by immunoprecipitation from HeLa cell nuclear extracts. In vitro transcription from the PAI-1 promoter in HeLa cell extracts was inhibited by HLTF antibodies and by the HLTF DNA binding domain. Over-expression of HLTF or HLTFMet123 produced a three-fold induction of PAI-1-LUC transient expression in HeLa cells. Mutation of the PAI-1 B Box led to an eight-fold reduction of basal PAI-1-LUC expression in these cell lines, but did not affect the four- to six-fold induction by phorbol esters.
Ding
H
H
Center for Molecular and Vascular Biology, University of Leuven, Belgium.
Descheemaeker
K
K
Marynen
P
P
Nelles
L
L
Carvalho
T
T
Carmo-Fonseca
M
M
Collen
D
D
Belayew
A
A
D000595:Amino Acid Sequence; D001483:Base Sequence; D002467:Cell Nucleus / Q000737:chemistry; D002893:Chromosomes, Human, Pair 3; D003001:Cloning, Molecular; D004247:DNA / Q000378:metabolism; D004265:DNA Helicases / Q000235:genetics; D018076:DNA, Complementary / Q000235:genetics; D004268:DNA-Binding Proteins / Q000032:analysis / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D005786:Gene Expression Regulation / Q000502:physiology*; D006367:HeLa Cells; D006801:Humans; D008969:Molecular Sequence Data; D008970:Molecular Weight; D018482:Muscle, Skeletal; D009928:Organ Specificity; D017395:Plasminogen Activator Inhibitor 1 / Q000235:genetics*; D011485:Protein Binding; D012333:RNA, Messenger / Q000032:analysis; D012045:Regulatory Sequences, Nucleic Acid / Q000235:genetics; D017422:Sequence Analysis, DNA; D014157:Transcription Factors / Q000032:analysis / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D014158:Transcription, Genetic / Q000235:genetics; D016335:Zinc Fingers / Q000235:genetics
D018076:DNA, Complementary; D004268:DNA-Binding Proteins; C100023:HLTF protein, human; D017395:Plasminogen Activator Inhibitor 1; D012333:RNA, Messenger; D014157:Transcription Factors; D004247:DNA; D004265:DNA Helicases