Abstract
The major human AP-endonuclease 1 (APE1) is a multifunctional protein that plays a central role in the repair of damaged DNA by acting as a dual-function nuclease in the base excision repair pathway. This enzyme was also independently identified as a redox activator of AP-1 DNA-binding activity and has subsequently been shown to activate a variety of transcription factors via a redox mechanism. In a third distinct role, APE1 was identified as a component of a trans-acting complex that acts as a repressor by binding to the negative calcium responsive elements (nCaRE)-A and nCaRE-B, which were first discovered in the promoter of the human parathyroid gene and later in the APE1 promoter itself. Here we show that the nuclear protein complex which binds to the nCaRE-B2 of the hAPE1 gene contains APE1 itself and the heterogeneous nuclear ribonucleoprotein L (hnRNP-L). The interaction between the APE1 and hnRNP-L proteins does not require the presence of nCaRE-B2. Our results support the possibility that the APE1 gene is down-regulated by its own product, which would be the first such example of the regulation of a DNA repair enzyme, and identify a novel function of hnRNP-L in transcriptional regulation.
Publication types
-
Research Support, U.S. Gov't, P.H.S.
MeSH terms
-
Amino Acid Sequence
-
Animals
-
Base Sequence
-
COS Cells
-
Calcium / pharmacology*
-
Carbon-Oxygen Lyases / genetics*
-
Carbon-Oxygen Lyases / isolation & purification
-
Carbon-Oxygen Lyases / metabolism*
-
Chromatography, Affinity
-
DNA / genetics
-
DNA / metabolism
-
DNA-(Apurinic or Apyrimidinic Site) Lyase
-
DNA-Binding Proteins / genetics
-
DNA-Binding Proteins / isolation & purification
-
DNA-Binding Proteins / metabolism
-
Down-Regulation
-
Electrophoretic Mobility Shift Assay
-
Feedback, Physiological
-
Gene Expression Regulation / drug effects
-
HeLa Cells
-
Heterogeneous-Nuclear Ribonucleoprotein L
-
Heterogeneous-Nuclear Ribonucleoproteins
-
Humans
-
Macromolecular Substances
-
Mutation / genetics
-
Promoter Regions, Genetic / genetics*
-
Protein Binding
-
Repressor Proteins / genetics
-
Repressor Proteins / isolation & purification
-
Repressor Proteins / metabolism*
-
Response Elements / genetics*
-
Ribonucleoproteins / chemistry
-
Ribonucleoproteins / genetics
-
Ribonucleoproteins / isolation & purification
-
Ribonucleoproteins / metabolism*
-
Sequence Alignment
-
Sequence Analysis, Protein
-
Transcription, Genetic / drug effects
Substances
-
DNA-Binding Proteins
-
Heterogeneous-Nuclear Ribonucleoprotein L
-
Heterogeneous-Nuclear Ribonucleoproteins
-
Macromolecular Substances
-
Repressor Proteins
-
Ribonucleoproteins
-
DNA
-
Carbon-Oxygen Lyases
-
APEX1 protein, human
-
DNA-(Apurinic or Apyrimidinic Site) Lyase
-
Calcium