Cloning Two Isoforms of Rat Cyclooxygenase

Differential Regulation of Their Expression

L. Feng, W. Q. Sun, Y. Y. Xia, W. W. Tang, P. Chanmugam, E. Soyoola, C. B. Wilson, D. Hwang

Research output: Contribution to journalArticle

443 Citations (Scopus)

Abstract

Two isoforms of cyclooxygenase (COX) have been identified in eukaryotic cells: COX-1 encoded by a 2.8-kb mRNA, and a mitogen-inducible COX-2 encoded by a 4-kb mRNA. We have cloned the COX-1 and COX-2 cDNAs from the cDNA library constructed from lipopolysaccharide (LPS)-stimulated rat peritoneal macrophages. The deduced amino acid sequence showed that COX- I contained 602 amino acids, whereas COX-2 contained 604 amino acids. There is 95% conservation of the nucleotide sequence in the open reading frame of COX-1 between the rat and the mouse, while the homology of the 3′ untranslated region is 68% except for a 150 bp segment adjacent to the stop codon which is nonhomologous with the mouse. Transfection of both COX cDNAs into Cos-7 cells resulted in increased COX activity. In rat vascular smooth muscle cells, interleukin-1β selectively increased the expression of COX-2, but not that of COX-1, as assessed by enzyme activity, immunoprecipitation of COX proteins, and mRNA analysis. Only the brain among tissues tested exhibits basal expression of COX-2 as the major form of the enzyme. However, COX-2 mRNA was expressed in vivo in the lung and kidney, but not in the heart, after systemic administration of LPS, suggesting that COX-2 but not COX-1 plays a major role in producing COX-derived products of arachidonic acid during endotoxic shock. Thus, the two COX isoforms were differentially expressed, and COX-2 was selectively induced in response to inflammatory stimuli in rats.

Original languageEnglish (US)
Pages (from-to)361-368
Number of pages8
JournalArchives of Biochemistry and Biophysics
Volume307
Issue number2
DOIs
StatePublished - Dec 1993
Externally publishedYes

Fingerprint

Cloning
Cyclooxygenase 2
Prostaglandin-Endoperoxide Synthases
Rats
Organism Cloning
Protein Isoforms
Cyclooxygenase 1
Messenger RNA
Amino Acids
Lipopolysaccharides
Complementary DNA
Terminator Codon
Macrophages
Peritoneal Macrophages
Enzyme activity
3' Untranslated Regions
Eukaryotic Cells
Enzymes
Septic Shock
Interleukin-1

ASJC Scopus subject areas

  • Molecular Biology
  • Biophysics
  • Biochemistry

Cite this

Feng, L., Sun, W. Q., Xia, Y. Y., Tang, W. W., Chanmugam, P., Soyoola, E., ... Hwang, D. (1993). Cloning Two Isoforms of Rat Cyclooxygenase: Differential Regulation of Their Expression. Archives of Biochemistry and Biophysics, 307(2), 361-368. https://doi.org/10.1006/abbi.1993.1601

Cloning Two Isoforms of Rat Cyclooxygenase : Differential Regulation of Their Expression. / Feng, L.; Sun, W. Q.; Xia, Y. Y.; Tang, W. W.; Chanmugam, P.; Soyoola, E.; Wilson, C. B.; Hwang, D.

In: Archives of Biochemistry and Biophysics, Vol. 307, No. 2, 12.1993, p. 361-368.

Research output: Contribution to journalArticle

Feng, L, Sun, WQ, Xia, YY, Tang, WW, Chanmugam, P, Soyoola, E, Wilson, CB & Hwang, D 1993, 'Cloning Two Isoforms of Rat Cyclooxygenase: Differential Regulation of Their Expression', Archives of Biochemistry and Biophysics, vol. 307, no. 2, pp. 361-368. https://doi.org/10.1006/abbi.1993.1601
Feng, L. ; Sun, W. Q. ; Xia, Y. Y. ; Tang, W. W. ; Chanmugam, P. ; Soyoola, E. ; Wilson, C. B. ; Hwang, D. / Cloning Two Isoforms of Rat Cyclooxygenase : Differential Regulation of Their Expression. In: Archives of Biochemistry and Biophysics. 1993 ; Vol. 307, No. 2. pp. 361-368.
@article{7b4e6f2344be415bbc0dcb90b5515135,
title = "Cloning Two Isoforms of Rat Cyclooxygenase: Differential Regulation of Their Expression",
abstract = "Two isoforms of cyclooxygenase (COX) have been identified in eukaryotic cells: COX-1 encoded by a 2.8-kb mRNA, and a mitogen-inducible COX-2 encoded by a 4-kb mRNA. We have cloned the COX-1 and COX-2 cDNAs from the cDNA library constructed from lipopolysaccharide (LPS)-stimulated rat peritoneal macrophages. The deduced amino acid sequence showed that COX- I contained 602 amino acids, whereas COX-2 contained 604 amino acids. There is 95{\%} conservation of the nucleotide sequence in the open reading frame of COX-1 between the rat and the mouse, while the homology of the 3′ untranslated region is 68{\%} except for a 150 bp segment adjacent to the stop codon which is nonhomologous with the mouse. Transfection of both COX cDNAs into Cos-7 cells resulted in increased COX activity. In rat vascular smooth muscle cells, interleukin-1β selectively increased the expression of COX-2, but not that of COX-1, as assessed by enzyme activity, immunoprecipitation of COX proteins, and mRNA analysis. Only the brain among tissues tested exhibits basal expression of COX-2 as the major form of the enzyme. However, COX-2 mRNA was expressed in vivo in the lung and kidney, but not in the heart, after systemic administration of LPS, suggesting that COX-2 but not COX-1 plays a major role in producing COX-derived products of arachidonic acid during endotoxic shock. Thus, the two COX isoforms were differentially expressed, and COX-2 was selectively induced in response to inflammatory stimuli in rats.",
author = "L. Feng and Sun, {W. Q.} and Xia, {Y. Y.} and Tang, {W. W.} and P. Chanmugam and E. Soyoola and Wilson, {C. B.} and D. Hwang",
year = "1993",
month = "12",
doi = "10.1006/abbi.1993.1601",
language = "English (US)",
volume = "307",
pages = "361--368",
journal = "Archives of Biochemistry and Biophysics",
issn = "0003-9861",
publisher = "Academic Press Inc.",
number = "2",

}

TY - JOUR

T1 - Cloning Two Isoforms of Rat Cyclooxygenase

T2 - Differential Regulation of Their Expression

AU - Feng, L.

AU - Sun, W. Q.

AU - Xia, Y. Y.

AU - Tang, W. W.

AU - Chanmugam, P.

AU - Soyoola, E.

AU - Wilson, C. B.

AU - Hwang, D.

PY - 1993/12

Y1 - 1993/12

N2 - Two isoforms of cyclooxygenase (COX) have been identified in eukaryotic cells: COX-1 encoded by a 2.8-kb mRNA, and a mitogen-inducible COX-2 encoded by a 4-kb mRNA. We have cloned the COX-1 and COX-2 cDNAs from the cDNA library constructed from lipopolysaccharide (LPS)-stimulated rat peritoneal macrophages. The deduced amino acid sequence showed that COX- I contained 602 amino acids, whereas COX-2 contained 604 amino acids. There is 95% conservation of the nucleotide sequence in the open reading frame of COX-1 between the rat and the mouse, while the homology of the 3′ untranslated region is 68% except for a 150 bp segment adjacent to the stop codon which is nonhomologous with the mouse. Transfection of both COX cDNAs into Cos-7 cells resulted in increased COX activity. In rat vascular smooth muscle cells, interleukin-1β selectively increased the expression of COX-2, but not that of COX-1, as assessed by enzyme activity, immunoprecipitation of COX proteins, and mRNA analysis. Only the brain among tissues tested exhibits basal expression of COX-2 as the major form of the enzyme. However, COX-2 mRNA was expressed in vivo in the lung and kidney, but not in the heart, after systemic administration of LPS, suggesting that COX-2 but not COX-1 plays a major role in producing COX-derived products of arachidonic acid during endotoxic shock. Thus, the two COX isoforms were differentially expressed, and COX-2 was selectively induced in response to inflammatory stimuli in rats.

AB - Two isoforms of cyclooxygenase (COX) have been identified in eukaryotic cells: COX-1 encoded by a 2.8-kb mRNA, and a mitogen-inducible COX-2 encoded by a 4-kb mRNA. We have cloned the COX-1 and COX-2 cDNAs from the cDNA library constructed from lipopolysaccharide (LPS)-stimulated rat peritoneal macrophages. The deduced amino acid sequence showed that COX- I contained 602 amino acids, whereas COX-2 contained 604 amino acids. There is 95% conservation of the nucleotide sequence in the open reading frame of COX-1 between the rat and the mouse, while the homology of the 3′ untranslated region is 68% except for a 150 bp segment adjacent to the stop codon which is nonhomologous with the mouse. Transfection of both COX cDNAs into Cos-7 cells resulted in increased COX activity. In rat vascular smooth muscle cells, interleukin-1β selectively increased the expression of COX-2, but not that of COX-1, as assessed by enzyme activity, immunoprecipitation of COX proteins, and mRNA analysis. Only the brain among tissues tested exhibits basal expression of COX-2 as the major form of the enzyme. However, COX-2 mRNA was expressed in vivo in the lung and kidney, but not in the heart, after systemic administration of LPS, suggesting that COX-2 but not COX-1 plays a major role in producing COX-derived products of arachidonic acid during endotoxic shock. Thus, the two COX isoforms were differentially expressed, and COX-2 was selectively induced in response to inflammatory stimuli in rats.

UR - http://www.scopus.com/inward/record.url?scp=0027141004&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0027141004&partnerID=8YFLogxK

U2 - 10.1006/abbi.1993.1601

DO - 10.1006/abbi.1993.1601

M3 - Article

VL - 307

SP - 361

EP - 368

JO - Archives of Biochemistry and Biophysics

JF - Archives of Biochemistry and Biophysics

SN - 0003-9861

IS - 2

ER -