The HIP1 Binding Site Is Required for Growth Regulation of the Dihydrofolate Reductase Gene Promoter

Anna L. Means, Jill E. Slansky, Stephanie L. McMahon, Mark W. Knuth, Peggy J. Farnham

Research output: Contribution to journalArticle

127 Citations (Scopus)

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.

Original languageEnglish (US)
Pages (from-to)1054-1063
Number of pages10
JournalMolecular and Cellular Biology
Volume12
Issue number3
StatePublished - Mar 1992
Externally publishedYes

Fingerprint

Tetrahydrofolate Dehydrogenase
Binding Sites
Growth
Genes
NIH 3T3 Cells
Proteins
Reporter Genes
Protein Binding
Cell Cycle
Peptides
Mutation
DNA

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics
  • Cell Biology

Cite this

Means, A. L., Slansky, J. E., McMahon, S. L., Knuth, M. W., & Farnham, P. J. (1992). The HIP1 Binding Site Is Required for Growth Regulation of the Dihydrofolate Reductase Gene Promoter. Molecular and Cellular Biology, 12(3), 1054-1063.

The HIP1 Binding Site Is Required for Growth Regulation of the Dihydrofolate Reductase Gene Promoter. / Means, Anna L.; Slansky, Jill E.; McMahon, Stephanie L.; Knuth, Mark W.; Farnham, Peggy J.

In: Molecular and Cellular Biology, Vol. 12, No. 3, 03.1992, p. 1054-1063.

Research output: Contribution to journalArticle

Means, AL, Slansky, JE, McMahon, SL, Knuth, MW & Farnham, PJ 1992, 'The HIP1 Binding Site Is Required for Growth Regulation of the Dihydrofolate Reductase Gene Promoter', Molecular and Cellular Biology, vol. 12, no. 3, pp. 1054-1063.
Means, Anna L. ; Slansky, Jill E. ; McMahon, Stephanie L. ; Knuth, Mark W. ; Farnham, Peggy J. / The HIP1 Binding Site Is Required for Growth Regulation of the Dihydrofolate Reductase Gene Promoter. In: Molecular and Cellular Biology. 1992 ; Vol. 12, No. 3. pp. 1054-1063.
@article{1fd65a5e8c724b448b4eab0f8c6a3098,
title = "The HIP1 Binding Site Is Required for Growth Regulation of the Dihydrofolate Reductase Gene Promoter",
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.",
author = "Means, {Anna L.} and Slansky, {Jill E.} and McMahon, {Stephanie L.} and Knuth, {Mark W.} and Farnham, {Peggy J.}",
year = "1992",
month = "3",
language = "English (US)",
volume = "12",
pages = "1054--1063",
journal = "Molecular and Cellular Biology",
issn = "0270-7306",
publisher = "American Society for Microbiology",
number = "3",

}

TY - JOUR

T1 - The HIP1 Binding Site Is Required for Growth Regulation of the Dihydrofolate Reductase Gene Promoter

AU - Means, Anna L.

AU - Slansky, Jill E.

AU - McMahon, Stephanie L.

AU - Knuth, Mark W.

AU - Farnham, Peggy J.

PY - 1992/3

Y1 - 1992/3

N2 - 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.

AB - 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.

UR - http://www.scopus.com/inward/record.url?scp=0026562757&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0026562757&partnerID=8YFLogxK

M3 - Article

C2 - 1545788

AN - SCOPUS:0026562757

VL - 12

SP - 1054

EP - 1063

JO - Molecular and Cellular Biology

JF - Molecular and Cellular Biology

SN - 0270-7306

IS - 3

ER -