Naturally occurring mutations in the human 5-lipoxygenase gene promoter that modify transcription factor binding and reporter gene transcription

Kwang Ho In, K. Asano, D. Beier, J. Grobholz, P. W. Finn, E. K. Silverman, E. S. Silverman, T. Collins, A. R. Fischer, T. P. Keith, K. Serino, S. W. Kim, G. T. De Sanctis, C. Yandava, A. Pillari, P. Rubin, J. Kemp, E. Israel, W. Busse, D. LedfordJ. J. Murray, A. Segal, D. Tinkleman, J. M. Drazen

Research output: Contribution to journalArticle

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Abstract

Five lipoxygenase (5-LO) is the first committed enzyme in the metabolic pathway leading to the synthesis of the leukotrienes. We examined genomic DNA isolated from 25 normal subjects and 31 patients with asthma (6 of whom had aspirin-sensitive asthma) for mutations in the known transcription factor binding regions and the protein encoding region of the 5-LO gene. A family of mutations in the G + C-rich transcription factor binding region was identified consisting of the deletion of one, deletion of two, or addition of one zinc finger (Sp1/Egr-1) binding sites in the region 176 to 147 bp upstream from the ATG translation start site where there are normally 5 Sp1 binding motifs in tandem. Reporter gene activity directed by any of the mutant forms of the transcription factor binding region was significantly (P < 0.05) less effective than the activity driven by the wild type transcription factor binding region. Electrophoretic mobility shift assays (EMSAs) demonstrated the capacity of wild type and mutant transcription factor binding regions to bind nuclear extracts from human umbilical vein endothelial cells (HUVECs). These data are consistent with a family of mutations in the 5-LO gene that can modify reporter gene transcription possibly through differences in Sp1 and Egr-1 transactivation.

Original languageEnglish
Pages (from-to)1130-1137
Number of pages8
JournalJournal of Clinical Investigation
Volume99
Issue number5
DOIs
Publication statusPublished - 1997 Mar 1
Externally publishedYes

Fingerprint

Arachidonate 5-Lipoxygenase
Reporter Genes
Transcription Factors
Mutation
Genes
Asthma
Leukotrienes
Zinc Fingers
Human Umbilical Vein Endothelial Cells
Electrophoretic Mobility Shift Assay
Metabolic Networks and Pathways
Transcriptional Activation
Aspirin
Carrier Proteins
Binding Sites
DNA
Enzymes

Keywords

  • asthma
  • leukotrienes

ASJC Scopus subject areas

  • Medicine(all)

Cite this

Naturally occurring mutations in the human 5-lipoxygenase gene promoter that modify transcription factor binding and reporter gene transcription. / In, Kwang Ho; Asano, K.; Beier, D.; Grobholz, J.; Finn, P. W.; Silverman, E. K.; Silverman, E. S.; Collins, T.; Fischer, A. R.; Keith, T. P.; Serino, K.; Kim, S. W.; De Sanctis, G. T.; Yandava, C.; Pillari, A.; Rubin, P.; Kemp, J.; Israel, E.; Busse, W.; Ledford, D.; Murray, J. J.; Segal, A.; Tinkleman, D.; Drazen, J. M.

In: Journal of Clinical Investigation, Vol. 99, No. 5, 01.03.1997, p. 1130-1137.

Research output: Contribution to journalArticle

In, KH, Asano, K, Beier, D, Grobholz, J, Finn, PW, Silverman, EK, Silverman, ES, Collins, T, Fischer, AR, Keith, TP, Serino, K, Kim, SW, De Sanctis, GT, Yandava, C, Pillari, A, Rubin, P, Kemp, J, Israel, E, Busse, W, Ledford, D, Murray, JJ, Segal, A, Tinkleman, D & Drazen, JM 1997, 'Naturally occurring mutations in the human 5-lipoxygenase gene promoter that modify transcription factor binding and reporter gene transcription', Journal of Clinical Investigation, vol. 99, no. 5, pp. 1130-1137. https://doi.org/10.1172/JCI119241
In, Kwang Ho ; Asano, K. ; Beier, D. ; Grobholz, J. ; Finn, P. W. ; Silverman, E. K. ; Silverman, E. S. ; Collins, T. ; Fischer, A. R. ; Keith, T. P. ; Serino, K. ; Kim, S. W. ; De Sanctis, G. T. ; Yandava, C. ; Pillari, A. ; Rubin, P. ; Kemp, J. ; Israel, E. ; Busse, W. ; Ledford, D. ; Murray, J. J. ; Segal, A. ; Tinkleman, D. ; Drazen, J. M. / Naturally occurring mutations in the human 5-lipoxygenase gene promoter that modify transcription factor binding and reporter gene transcription. In: Journal of Clinical Investigation. 1997 ; Vol. 99, No. 5. pp. 1130-1137.
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abstract = "Five lipoxygenase (5-LO) is the first committed enzyme in the metabolic pathway leading to the synthesis of the leukotrienes. We examined genomic DNA isolated from 25 normal subjects and 31 patients with asthma (6 of whom had aspirin-sensitive asthma) for mutations in the known transcription factor binding regions and the protein encoding region of the 5-LO gene. A family of mutations in the G + C-rich transcription factor binding region was identified consisting of the deletion of one, deletion of two, or addition of one zinc finger (Sp1/Egr-1) binding sites in the region 176 to 147 bp upstream from the ATG translation start site where there are normally 5 Sp1 binding motifs in tandem. Reporter gene activity directed by any of the mutant forms of the transcription factor binding region was significantly (P < 0.05) less effective than the activity driven by the wild type transcription factor binding region. Electrophoretic mobility shift assays (EMSAs) demonstrated the capacity of wild type and mutant transcription factor binding regions to bind nuclear extracts from human umbilical vein endothelial cells (HUVECs). These data are consistent with a family of mutations in the 5-LO gene that can modify reporter gene transcription possibly through differences in Sp1 and Egr-1 transactivation.",
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AU - In, Kwang Ho

AU - Asano, K.

AU - Beier, D.

AU - Grobholz, J.

AU - Finn, P. W.

AU - Silverman, E. K.

AU - Silverman, E. S.

AU - Collins, T.

AU - Fischer, A. R.

AU - Keith, T. P.

AU - Serino, K.

AU - Kim, S. W.

AU - De Sanctis, G. T.

AU - Yandava, C.

AU - Pillari, A.

AU - Rubin, P.

AU - Kemp, J.

AU - Israel, E.

AU - Busse, W.

AU - Ledford, D.

AU - Murray, J. J.

AU - Segal, A.

AU - Tinkleman, D.

AU - Drazen, J. M.

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N2 - Five lipoxygenase (5-LO) is the first committed enzyme in the metabolic pathway leading to the synthesis of the leukotrienes. We examined genomic DNA isolated from 25 normal subjects and 31 patients with asthma (6 of whom had aspirin-sensitive asthma) for mutations in the known transcription factor binding regions and the protein encoding region of the 5-LO gene. A family of mutations in the G + C-rich transcription factor binding region was identified consisting of the deletion of one, deletion of two, or addition of one zinc finger (Sp1/Egr-1) binding sites in the region 176 to 147 bp upstream from the ATG translation start site where there are normally 5 Sp1 binding motifs in tandem. Reporter gene activity directed by any of the mutant forms of the transcription factor binding region was significantly (P < 0.05) less effective than the activity driven by the wild type transcription factor binding region. Electrophoretic mobility shift assays (EMSAs) demonstrated the capacity of wild type and mutant transcription factor binding regions to bind nuclear extracts from human umbilical vein endothelial cells (HUVECs). These data are consistent with a family of mutations in the 5-LO gene that can modify reporter gene transcription possibly through differences in Sp1 and Egr-1 transactivation.

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