Hepatic nuclear receptor PPAR in the koala (Phascolarctos cinereus)

Cloning and molecular characterisation

Suong Ngo, R MCKINNON, I Stupans

    Research output: Contribution to journalArticleResearchpeer-review

    Abstract

    Peroxisome proliferator-activated receptor ? (PPAR?) is a member of the nuclear/steroid receptor gene superfamily that plays an essential role in fatty acid metabolism. PPAR? modulates the expression of genes encoding peroxisomal fatty acid ?-oxidation enzymes and microsomal fatty acid hydroxylases CYP4As. We have previously reported that the obligate Eucalyptus feeder koala (Phascolarctos cinereus) exhibits a higher hepatic CYP4A activity and an absence of peroxisomal palmitoyl-CoA oxidation as compared to non-Eucalyptus feeders human, rat or wallaby. Here we describe the cloning, expression and molecular characterisation of koala hepatic PPAR?. A full-length PPAR? cDNA of size 1515�bp was cloned by reverse transcription-polymerase chain reaction (RT-PCR) and rapid amplification of cDNA ends (RACE). The koala PPAR? cDNA encodes a protein of 468 amino acids. Transfection of the koala PPAR? cDNA into Cos-7 cells resulted in the expression of a protein recognised by a rabbit anti-human PPAR? polyclonal antibody. PPAR? immunoreactive bands of the same molecular mass were detected in nuclear extracts of koala livers. The results of this study demonstrate the presence of koala hepatic PPAR? which shares several common features with other published PPAR?s; however, it exhibits important differences in both the DNA and ligand binding domains. � 2007 Elsevier Inc. All rights reserved.
    Original languageEnglish
    Pages (from-to)375-382
    Number of pages8
    JournalComparative Biochemistry and Physiology, Part C
    Volume146
    Issue number3
    Publication statusPublished - 2007

    Fingerprint

    Phascolarctidae
    Peroxisome Proliferator-Activated Receptors
    Cloning
    Molecular Cloning
    Cytoplasmic and Nuclear Receptors
    Liver
    Complementary DNA
    Cytochrome P-450 CYP4A
    Fatty Acids
    Palmitoyl Coenzyme A
    Macropodidae
    Liver Extracts
    Eucalyptus
    Oxidation
    Gene encoding
    Steroid Receptors
    Polymerase chain reaction
    Molecular mass
    Transcription
    Mixed Function Oxygenases

    Cite this

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    abstract = "Peroxisome proliferator-activated receptor ? (PPAR?) is a member of the nuclear/steroid receptor gene superfamily that plays an essential role in fatty acid metabolism. PPAR? modulates the expression of genes encoding peroxisomal fatty acid ?-oxidation enzymes and microsomal fatty acid hydroxylases CYP4As. We have previously reported that the obligate Eucalyptus feeder koala (Phascolarctos cinereus) exhibits a higher hepatic CYP4A activity and an absence of peroxisomal palmitoyl-CoA oxidation as compared to non-Eucalyptus feeders human, rat or wallaby. Here we describe the cloning, expression and molecular characterisation of koala hepatic PPAR?. A full-length PPAR? cDNA of size 1515�bp was cloned by reverse transcription-polymerase chain reaction (RT-PCR) and rapid amplification of cDNA ends (RACE). The koala PPAR? cDNA encodes a protein of 468 amino acids. Transfection of the koala PPAR? cDNA into Cos-7 cells resulted in the expression of a protein recognised by a rabbit anti-human PPAR? polyclonal antibody. PPAR? immunoreactive bands of the same molecular mass were detected in nuclear extracts of koala livers. The results of this study demonstrate the presence of koala hepatic PPAR? which shares several common features with other published PPAR?s; however, it exhibits important differences in both the DNA and ligand binding domains. � 2007 Elsevier Inc. All rights reserved.",
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    author = "Suong Ngo and R MCKINNON and I Stupans",
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    Hepatic nuclear receptor PPAR in the koala (Phascolarctos cinereus) : Cloning and molecular characterisation. / Ngo, Suong; MCKINNON, R; Stupans, I.

    In: Comparative Biochemistry and Physiology, Part C, Vol. 146, No. 3, 2007, p. 375-382.

    Research output: Contribution to journalArticleResearchpeer-review

    TY - JOUR

    T1 - Hepatic nuclear receptor PPAR in the koala (Phascolarctos cinereus)

    T2 - Cloning and molecular characterisation

    AU - Ngo, Suong

    AU - MCKINNON, R

    AU - Stupans, I

    PY - 2007

    Y1 - 2007

    N2 - Peroxisome proliferator-activated receptor ? (PPAR?) is a member of the nuclear/steroid receptor gene superfamily that plays an essential role in fatty acid metabolism. PPAR? modulates the expression of genes encoding peroxisomal fatty acid ?-oxidation enzymes and microsomal fatty acid hydroxylases CYP4As. We have previously reported that the obligate Eucalyptus feeder koala (Phascolarctos cinereus) exhibits a higher hepatic CYP4A activity and an absence of peroxisomal palmitoyl-CoA oxidation as compared to non-Eucalyptus feeders human, rat or wallaby. Here we describe the cloning, expression and molecular characterisation of koala hepatic PPAR?. A full-length PPAR? cDNA of size 1515�bp was cloned by reverse transcription-polymerase chain reaction (RT-PCR) and rapid amplification of cDNA ends (RACE). The koala PPAR? cDNA encodes a protein of 468 amino acids. Transfection of the koala PPAR? cDNA into Cos-7 cells resulted in the expression of a protein recognised by a rabbit anti-human PPAR? polyclonal antibody. PPAR? immunoreactive bands of the same molecular mass were detected in nuclear extracts of koala livers. The results of this study demonstrate the presence of koala hepatic PPAR? which shares several common features with other published PPAR?s; however, it exhibits important differences in both the DNA and ligand binding domains. � 2007 Elsevier Inc. All rights reserved.

    AB - Peroxisome proliferator-activated receptor ? (PPAR?) is a member of the nuclear/steroid receptor gene superfamily that plays an essential role in fatty acid metabolism. PPAR? modulates the expression of genes encoding peroxisomal fatty acid ?-oxidation enzymes and microsomal fatty acid hydroxylases CYP4As. We have previously reported that the obligate Eucalyptus feeder koala (Phascolarctos cinereus) exhibits a higher hepatic CYP4A activity and an absence of peroxisomal palmitoyl-CoA oxidation as compared to non-Eucalyptus feeders human, rat or wallaby. Here we describe the cloning, expression and molecular characterisation of koala hepatic PPAR?. A full-length PPAR? cDNA of size 1515�bp was cloned by reverse transcription-polymerase chain reaction (RT-PCR) and rapid amplification of cDNA ends (RACE). The koala PPAR? cDNA encodes a protein of 468 amino acids. Transfection of the koala PPAR? cDNA into Cos-7 cells resulted in the expression of a protein recognised by a rabbit anti-human PPAR? polyclonal antibody. PPAR? immunoreactive bands of the same molecular mass were detected in nuclear extracts of koala livers. The results of this study demonstrate the presence of koala hepatic PPAR? which shares several common features with other published PPAR?s; however, it exhibits important differences in both the DNA and ligand binding domains. � 2007 Elsevier Inc. All rights reserved.

    KW - complementary DNA

    KW - cytochrome P450 4A

    KW - palmitoyl coenzyme A

    KW - peroxisome proliferator activated receptor alpha

    KW - animal cell

    KW - animal tissue

    KW - article

    KW - cell strain COS7

    KW - controlled study

    KW - DNA binding

    KW - enzyme activity

    KW - Eucalyptus

    KW - fatty acid oxidation

    KW - koala

    KW - ligand binding

    KW - liver metabolism

    KW - male

    KW - molecular cloning

    KW - nonhuman

    KW - nucleotide sequence

    KW - priority journal

    KW - protein domain

    KW - reverse transcription polymerase chain reaction

    KW - sequence homology

    KW - Amino Acid Sequence

    KW - Animals

    KW - Base Sequence

    KW - Cercopithecus aethiops

    KW - Cloning, Molecular

    KW - COS Cells

    KW - Humans

    KW - Liver

    KW - Male

    KW - Mice

    KW - Molecular Sequence Data

    KW - Phascolarctidae

    KW - PPAR alpha

    KW - Rats

    KW - Reverse Transcriptase Polymerase Chain Reaction

    KW - RNA, Messenger

    KW - Sequence Analysis, Protein

    KW - Species Specificity

    KW - Oryctolagus cuniculus

    KW - Phascolarctos cinereus

    KW - Rattus

    M3 - Article

    VL - 146

    SP - 375

    EP - 382

    JO - Comparative Biochemistry and Physiology, Part C

    JF - Comparative Biochemistry and Physiology, Part C

    SN - 1532-0456

    IS - 3

    ER -