Analysis of the rat lactate dehydrogenase A subunit gene promoter/regulatory region

M. L. Short, D. Huang, D. M. Milkowski, S. Short, K. Kunstman, C. J. Soong, K. C. Chung, R. A. Jungmann

Research output: Contribution to journalArticle

39 Citations (Scopus)

Abstract

The rat lactate dehydrogenase (LDH) A subunit gene promoter contains a putative AP-1 binding site at -295/-289 bp, two consensus Sp1 binding sites at -141/-136 bp and -103/-98 bp, and a single copy of a consensus cyclic AMP-responsive element (CRE) at -48 to -41 bp upstream of the transcription initiation site. Additionally, an as yet unidentified silencer element is located within the -1173/-830 bp 5'-flanking region. Transient transfection analyses of a -1173/+25 bp LDH A-chloramphenicol acetyltransferase fusion gene has indicated a complete inability of the promoter fragment to direct basal or forskolin-induced transcription. Deletion of the -1173/-830 bp sequence restored basal and cyclic AMP (cAMP)-inducible activity. Point mutations in the Sp1 binding sites of a -830/+25 bp promoter fragment reduced basal but not the relative degree of cAMP-inducible activity. cAMP-regulated transcriptional activity was dependent upon an 8 bp CRE, -TGACGTCA-, located at the -48/-41 bp upstream region. Mutations in the CRE abolished cAMP-mediated induction and reduced basal activity by about 65%. The CRE binds a 47 kDa protein which has previously been identified as CRE binding protein (CREB)-327, an isoform of the activating transcription factor/CREB transcription factor gene family. Co-transfection of a vector that expresses the catalytic subunit of cAMP-dependent protein kinase stimulates LDH A subunit promoter activity suggesting that cAMP induces LDH A subunit gene expression through phosphorylative modification of CREB-327. This study emphasizes a fundamental role of several modules including Sp1 and CREB binding sites in regulating basal and cAMP-mediated transcriptional activity of the LDH A gene.

Original languageEnglish
Pages (from-to)391-398
Number of pages8
JournalBiochemical Journal
Volume304
Issue number2
DOIs
Publication statusPublished - 1994 Jan 1

Fingerprint

Nucleic Acid Regulatory Sequences
Genetic Promoter Regions
Cyclic AMP
Rats
Genes
Carrier Proteins
Binding Sites
Transfection
lactate dehydrogenase 5
Transcriptional Silencer Elements
Cyclic AMP-Dependent Protein Kinase Catalytic Subunits
Activating Transcription Factors
Chloramphenicol O-Acetyltransferase
5' Flanking Region
Transcription Initiation Site
Gene Fusion
Transcription Factor AP-1
Colforsin
Transcription
Point Mutation

All Science Journal Classification (ASJC) codes

  • Biochemistry
  • Molecular Biology
  • Cell Biology

Cite this

Short, M. L., Huang, D., Milkowski, D. M., Short, S., Kunstman, K., Soong, C. J., ... Jungmann, R. A. (1994). Analysis of the rat lactate dehydrogenase A subunit gene promoter/regulatory region. Biochemical Journal, 304(2), 391-398. https://doi.org/10.1042/bj3040391
Short, M. L. ; Huang, D. ; Milkowski, D. M. ; Short, S. ; Kunstman, K. ; Soong, C. J. ; Chung, K. C. ; Jungmann, R. A. / Analysis of the rat lactate dehydrogenase A subunit gene promoter/regulatory region. In: Biochemical Journal. 1994 ; Vol. 304, No. 2. pp. 391-398.
@article{4dca382b6be443f381df9788c17f7f1f,
title = "Analysis of the rat lactate dehydrogenase A subunit gene promoter/regulatory region",
abstract = "The rat lactate dehydrogenase (LDH) A subunit gene promoter contains a putative AP-1 binding site at -295/-289 bp, two consensus Sp1 binding sites at -141/-136 bp and -103/-98 bp, and a single copy of a consensus cyclic AMP-responsive element (CRE) at -48 to -41 bp upstream of the transcription initiation site. Additionally, an as yet unidentified silencer element is located within the -1173/-830 bp 5'-flanking region. Transient transfection analyses of a -1173/+25 bp LDH A-chloramphenicol acetyltransferase fusion gene has indicated a complete inability of the promoter fragment to direct basal or forskolin-induced transcription. Deletion of the -1173/-830 bp sequence restored basal and cyclic AMP (cAMP)-inducible activity. Point mutations in the Sp1 binding sites of a -830/+25 bp promoter fragment reduced basal but not the relative degree of cAMP-inducible activity. cAMP-regulated transcriptional activity was dependent upon an 8 bp CRE, -TGACGTCA-, located at the -48/-41 bp upstream region. Mutations in the CRE abolished cAMP-mediated induction and reduced basal activity by about 65{\%}. The CRE binds a 47 kDa protein which has previously been identified as CRE binding protein (CREB)-327, an isoform of the activating transcription factor/CREB transcription factor gene family. Co-transfection of a vector that expresses the catalytic subunit of cAMP-dependent protein kinase stimulates LDH A subunit promoter activity suggesting that cAMP induces LDH A subunit gene expression through phosphorylative modification of CREB-327. This study emphasizes a fundamental role of several modules including Sp1 and CREB binding sites in regulating basal and cAMP-mediated transcriptional activity of the LDH A gene.",
author = "Short, {M. L.} and D. Huang and Milkowski, {D. M.} and S. Short and K. Kunstman and Soong, {C. J.} and Chung, {K. C.} and Jungmann, {R. A.}",
year = "1994",
month = "1",
day = "1",
doi = "10.1042/bj3040391",
language = "English",
volume = "304",
pages = "391--398",
journal = "Biochemical Journal",
issn = "0264-6021",
publisher = "Portland Press Ltd.",
number = "2",

}

Short, ML, Huang, D, Milkowski, DM, Short, S, Kunstman, K, Soong, CJ, Chung, KC & Jungmann, RA 1994, 'Analysis of the rat lactate dehydrogenase A subunit gene promoter/regulatory region', Biochemical Journal, vol. 304, no. 2, pp. 391-398. https://doi.org/10.1042/bj3040391

Analysis of the rat lactate dehydrogenase A subunit gene promoter/regulatory region. / Short, M. L.; Huang, D.; Milkowski, D. M.; Short, S.; Kunstman, K.; Soong, C. J.; Chung, K. C.; Jungmann, R. A.

In: Biochemical Journal, Vol. 304, No. 2, 01.01.1994, p. 391-398.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Analysis of the rat lactate dehydrogenase A subunit gene promoter/regulatory region

AU - Short, M. L.

AU - Huang, D.

AU - Milkowski, D. M.

AU - Short, S.

AU - Kunstman, K.

AU - Soong, C. J.

AU - Chung, K. C.

AU - Jungmann, R. A.

PY - 1994/1/1

Y1 - 1994/1/1

N2 - The rat lactate dehydrogenase (LDH) A subunit gene promoter contains a putative AP-1 binding site at -295/-289 bp, two consensus Sp1 binding sites at -141/-136 bp and -103/-98 bp, and a single copy of a consensus cyclic AMP-responsive element (CRE) at -48 to -41 bp upstream of the transcription initiation site. Additionally, an as yet unidentified silencer element is located within the -1173/-830 bp 5'-flanking region. Transient transfection analyses of a -1173/+25 bp LDH A-chloramphenicol acetyltransferase fusion gene has indicated a complete inability of the promoter fragment to direct basal or forskolin-induced transcription. Deletion of the -1173/-830 bp sequence restored basal and cyclic AMP (cAMP)-inducible activity. Point mutations in the Sp1 binding sites of a -830/+25 bp promoter fragment reduced basal but not the relative degree of cAMP-inducible activity. cAMP-regulated transcriptional activity was dependent upon an 8 bp CRE, -TGACGTCA-, located at the -48/-41 bp upstream region. Mutations in the CRE abolished cAMP-mediated induction and reduced basal activity by about 65%. The CRE binds a 47 kDa protein which has previously been identified as CRE binding protein (CREB)-327, an isoform of the activating transcription factor/CREB transcription factor gene family. Co-transfection of a vector that expresses the catalytic subunit of cAMP-dependent protein kinase stimulates LDH A subunit promoter activity suggesting that cAMP induces LDH A subunit gene expression through phosphorylative modification of CREB-327. This study emphasizes a fundamental role of several modules including Sp1 and CREB binding sites in regulating basal and cAMP-mediated transcriptional activity of the LDH A gene.

AB - The rat lactate dehydrogenase (LDH) A subunit gene promoter contains a putative AP-1 binding site at -295/-289 bp, two consensus Sp1 binding sites at -141/-136 bp and -103/-98 bp, and a single copy of a consensus cyclic AMP-responsive element (CRE) at -48 to -41 bp upstream of the transcription initiation site. Additionally, an as yet unidentified silencer element is located within the -1173/-830 bp 5'-flanking region. Transient transfection analyses of a -1173/+25 bp LDH A-chloramphenicol acetyltransferase fusion gene has indicated a complete inability of the promoter fragment to direct basal or forskolin-induced transcription. Deletion of the -1173/-830 bp sequence restored basal and cyclic AMP (cAMP)-inducible activity. Point mutations in the Sp1 binding sites of a -830/+25 bp promoter fragment reduced basal but not the relative degree of cAMP-inducible activity. cAMP-regulated transcriptional activity was dependent upon an 8 bp CRE, -TGACGTCA-, located at the -48/-41 bp upstream region. Mutations in the CRE abolished cAMP-mediated induction and reduced basal activity by about 65%. The CRE binds a 47 kDa protein which has previously been identified as CRE binding protein (CREB)-327, an isoform of the activating transcription factor/CREB transcription factor gene family. Co-transfection of a vector that expresses the catalytic subunit of cAMP-dependent protein kinase stimulates LDH A subunit promoter activity suggesting that cAMP induces LDH A subunit gene expression through phosphorylative modification of CREB-327. This study emphasizes a fundamental role of several modules including Sp1 and CREB binding sites in regulating basal and cAMP-mediated transcriptional activity of the LDH A gene.

UR - http://www.scopus.com/inward/record.url?scp=0028099552&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0028099552&partnerID=8YFLogxK

U2 - 10.1042/bj3040391

DO - 10.1042/bj3040391

M3 - Article

C2 - 7998973

AN - SCOPUS:0028099552

VL - 304

SP - 391

EP - 398

JO - Biochemical Journal

JF - Biochemical Journal

SN - 0264-6021

IS - 2

ER -

Short ML, Huang D, Milkowski DM, Short S, Kunstman K, Soong CJ et al. Analysis of the rat lactate dehydrogenase A subunit gene promoter/regulatory region. Biochemical Journal. 1994 Jan 1;304(2):391-398. https://doi.org/10.1042/bj3040391