Isolation of promoter for cytosolic phospholipase A2 (cPLA2)

Agnes Tay; Peter Maxwell; Zhen Guo Li; Howard Goldberg; Karl Skorecki

doi:10.1016/0167-4781(94)90299-2

Isolation of promoter for cytosolic phospholipase A₂ (cPLA2)

Agnes Tay, Peter Maxwell, Zhen Guo Li, Howard Goldberg, Karl Skorecki

University of Toronto

Research output: Contribution to journal › Article › peer-review

55 Scopus citations

Abstract

Cytosolic phospholipase A₂ (cPLA2) releases arachidonic acid from membrane phospholipids and is believed to be the rate-limiting enzyme in the arachidonic acid pathway. We report herein the isolation of a 3 kb fragment of rodent genomic DNA containing part of the first intron, the first exon and 5′-flanking sequence. The start site of transcription was mapped by 5′-rapid amplification of cDNA ends and corroborated by ribonuclease protection assay. The gene has a TATAless promoter with no classical Sp1 binding sites or initiator element. A microsatellite series of CA repeats was noted in the 5′-flanking region of both the rodent and human promoters. Deletion constructs have been analysed for luciferase activity and confirmed promoter activity.

Original language	English
Pages (from-to)	345-347
Number of pages	3
Journal	Biochimica et Biophysica Acta - Gene Structure and Expression
Volume	1217
Issue number	3
DOIs	https://doi.org/10.1016/0167-4781(94)90299-2
State	Published - 6 Apr 1994
Externally published	Yes

Bibliographical note

Funding Information:
We thank Dr. John Knopf for cPLA2 cDNA plasmids and Dr. E. Chester Ridgway for pA3LUC expression vector. We also thank Drs. G. Mills, H. Elshotz and R. Sladek for helpful suggestions and encouragement and Dr. Catherine Duff for chromosomal mapping. A.T. was supported by a National University of Singapore Overseas Graduate Scholarship and P.M. by a Medical Research Council Fellowship. This study was supported by grants from the Medical Research Council, Kidney Foundation and National Cancer Institute of Canada.

Funding

We thank Dr. John Knopf for cPLA2 cDNA plasmids and Dr. E. Chester Ridgway for pA3LUC expression vector. We also thank Drs. G. Mills, H. Elshotz and R. Sladek for helpful suggestions and encouragement and Dr. Catherine Duff for chromosomal mapping. A.T. was supported by a National University of Singapore Overseas Graduate Scholarship and P.M. by a Medical Research Council Fellowship. This study was supported by grants from the Medical Research Council, Kidney Foundation and National Cancer Institute of Canada.

Funders	Funder number
National Cancer Institute of Canada
Kidney Foundation of Central Pennsylvania
Medical Research Council
National University of Singapore

Keywords

Arachidonic acid pathway
Cytosol
Nucleotide sequence
Phospholipase A
Promoter

Access to Document

10.1016/0167-4781(94)90299-2

Cite this

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abstract = "Cytosolic phospholipase A2 (cPLA2) releases arachidonic acid from membrane phospholipids and is believed to be the rate-limiting enzyme in the arachidonic acid pathway. We report herein the isolation of a 3 kb fragment of rodent genomic DNA containing part of the first intron, the first exon and 5′-flanking sequence. The start site of transcription was mapped by 5′-rapid amplification of cDNA ends and corroborated by ribonuclease protection assay. The gene has a TATAless promoter with no classical Sp1 binding sites or initiator element. A microsatellite series of CA repeats was noted in the 5′-flanking region of both the rodent and human promoters. Deletion constructs have been analysed for luciferase activity and confirmed promoter activity.",

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author = "Agnes Tay and Peter Maxwell and Li, {Zhen Guo} and Howard Goldberg and Karl Skorecki",

note = "Funding Information: We thank Dr. John Knopf for cPLA2 cDNA plasmids and Dr. E. Chester Ridgway for pA3LUC expression vector. We also thank Drs. G. Mills, H. Elshotz and R. Sladek for helpful suggestions and encouragement and Dr. Catherine Duff for chromosomal mapping. A.T. was supported by a National University of Singapore Overseas Graduate Scholarship and P.M. by a Medical Research Council Fellowship. This study was supported by grants from the Medical Research Council, Kidney Foundation and National Cancer Institute of Canada.",

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AU - Maxwell, Peter

AU - Li, Zhen Guo

AU - Goldberg, Howard

AU - Skorecki, Karl

N1 - Funding Information: We thank Dr. John Knopf for cPLA2 cDNA plasmids and Dr. E. Chester Ridgway for pA3LUC expression vector. We also thank Drs. G. Mills, H. Elshotz and R. Sladek for helpful suggestions and encouragement and Dr. Catherine Duff for chromosomal mapping. A.T. was supported by a National University of Singapore Overseas Graduate Scholarship and P.M. by a Medical Research Council Fellowship. This study was supported by grants from the Medical Research Council, Kidney Foundation and National Cancer Institute of Canada.

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N2 - Cytosolic phospholipase A2 (cPLA2) releases arachidonic acid from membrane phospholipids and is believed to be the rate-limiting enzyme in the arachidonic acid pathway. We report herein the isolation of a 3 kb fragment of rodent genomic DNA containing part of the first intron, the first exon and 5′-flanking sequence. The start site of transcription was mapped by 5′-rapid amplification of cDNA ends and corroborated by ribonuclease protection assay. The gene has a TATAless promoter with no classical Sp1 binding sites or initiator element. A microsatellite series of CA repeats was noted in the 5′-flanking region of both the rodent and human promoters. Deletion constructs have been analysed for luciferase activity and confirmed promoter activity.

AB - Cytosolic phospholipase A2 (cPLA2) releases arachidonic acid from membrane phospholipids and is believed to be the rate-limiting enzyme in the arachidonic acid pathway. We report herein the isolation of a 3 kb fragment of rodent genomic DNA containing part of the first intron, the first exon and 5′-flanking sequence. The start site of transcription was mapped by 5′-rapid amplification of cDNA ends and corroborated by ribonuclease protection assay. The gene has a TATAless promoter with no classical Sp1 binding sites or initiator element. A microsatellite series of CA repeats was noted in the 5′-flanking region of both the rodent and human promoters. Deletion constructs have been analysed for luciferase activity and confirmed promoter activity.

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KW - Nucleotide sequence

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