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The HIP1 binding site is required for growth regulation of the dihydrofolate reductase gene promoter
- Source :
- Molecular and Cellular Biology. 12:1054-1063
- Publication Year :
- 1992
- Publisher :
- Informa UK Limited, 1992.
-
Abstract
- The transcription rate of the dihydrofolate reductase (DHFR) gene increases at the G1/S boundary of the proliferative cell cycle. Through analysis of transiently and stably transfected NIH 3T3 cells, we have now demonstrated that DHFR promoter sequences extending from -270 to +20 are sufficient to confer similar regulation on a reporter gene. Mutation of a protein binding site that spans sequences from -16 to +11 in the DHFR promoter resulted in loss of the transcriptional increase at the G1/S boundary. Purification of an activity from HeLa nuclear extract that binds to this region enriched for a 180-kDa polypeptide (HIP1). Using this HIP1 preparation, we have identified specific positions within the binding site that are critical for efficient protein-DNA interactions. An analysis of association and dissociation rates suggests that bound HIP1 protein can exchange rapidly with free protein. This rapid exchange may facilitate the burst of transcriptional activity from the DHFR promoter at the G1/S boundary.
- Subjects :
- Molecular Sequence Data
Transfection
medicine.disease_cause
DNA-binding protein
Gene Expression Regulation, Enzymologic
Mice
chemistry.chemical_compound
Dihydrofolate reductase
Gene expression
medicine
Animals
Binding site
Promoter Regions, Genetic
Gene
Molecular Biology
Reporter gene
Mutation
Binding Sites
Base Sequence
biology
3T3 Cells
DNA
Cell Biology
Molecular biology
DNA-Binding Proteins
Molecular Weight
Tetrahydrofolate Dehydrogenase
chemistry
Mutagenesis, Site-Directed
biology.protein
Cell Division
Research Article
Subjects
Details
- ISSN :
- 10985549 and 02707306
- Volume :
- 12
- Database :
- OpenAIRE
- Journal :
- Molecular and Cellular Biology
- Accession number :
- edsair.doi.dedup.....82a18feaaaeffd697606eb03cf1d7056