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Subfamily III of mammalian oxysterol-binding protein (OSBP) homologues: the expression and intracellular localization of ORP3, ORP6, and ORP7

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Abstract

The human OSBP related protein (ORP) family consists of 12 members, which can be divided into six subfamilies based on the genomic organization and amino acid homology. Here we performed basic characterization of subfamily III, which consists of three members: ORP3, ORP6, and ORP7. According to cDNA hybridization, the three genes are expressed in a tissue-specific manner. While ORP3 mRNA is most abundant in kidney, lymph nodes, and thymus, ORP6 shows highest expression in brain and skeletal muscle, and ORP7 in the gastrointestinal tract. Using monospecific peptide antibodies, we confirmed the presence of the three proteins in human and mouse tissues. ORP6 gene expression was induced upon differentiation of F9 embryonic carcinoma cells into parietal endoderm, while ORP3 and ORP7 mRNA levels were unchanged. In the F9 cells, endogenous ORP6 associated predominantly with the nuclear envelope. When expressed from the cDNA in cultured cells, the three proteins were distributed between the cytosol and endoplasmic reticulum (ER) membranes, with a minor portion found at the plasma membrane. Experiments with truncated constructs showed that the N-terminal portion of the proteins, containing a pleckstrin homology (PH) domain, has markedly strong plasma membrane targeting specificity, while the C-terminal half remains largely cytosolic. The expression data demonstrates that ORP3, -6, and -7 are not merely redundant gene products but show marked quantitative differences in tissue expression, suggesting tissue-specific aspects in their function. The dual targeting of the proteins indicates a putative role in communication between the ER and the plasma membrane.

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Acknowledgements

We are grateful to Ulla Kiiski, Tuulikki Leinonen, Seija Puomilahti, and Pirjo Ranta for expert technical assistance, to Dr. Giulia Chinetti (Pasteur Institute, Lille, France) for the monocyte and macrophage protein specimens, and to Dr. Ilkka Reima for critically reading the manuscript. The KIAA0704 cDNA was kindly provided by Dr. Takahiro Nagase (Kazusa DNA Research Institute, Kisarazu, Chiba, Japan).

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  1. Sanna Lehtonen

    Present address: Department of Cellular and Molecular Medicine, University of California San Diego, 9500 Gilman Drive, La Jolla, CA 92093-0651, USA

Authors and Affiliations

  1. Department of Molecular Medicine, National Public Health Institute, Biomedicum, PO Box 104, 00251, Helsinki, Finland

    Markku Lehto & Vesa M. Olkkonen

  2. Department of Pathology, Helsinki University Central Hospital, Peijas Hospital, Sairaalakatu 1, 01400, Vantaa, Finland

    Jukka Tienari

  3. Department of Pathology, Haartman Institute and Helsinki University Central Hospital, University of Helsinki, PO Box 21, 00014, Finland

    Sanna Lehtonen & Eero Lehtonen

  4. Department of Cellular and Molecular Medicine, University of California San Diego, 9500 Gilman Drive, La Jolla, CA 92093-0651, USA

    Eero Lehtonen

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  1. Markku Lehto
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Correspondence to Vesa M. Olkkonen.

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This study was supported by the Clinical Research Fund of Helsinki University Central Hospital (J.T.), the Academy of Finland (grant 51883 to M.L.; grants 49987, 50641, and 54301 to V.M.O.), the Sigrid Juselius Foundation, and the Finnish Cultural Foundation

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Lehto, M., Tienari, J., Lehtonen, S. et al. Subfamily III of mammalian oxysterol-binding protein (OSBP) homologues: the expression and intracellular localization of ORP3, ORP6, and ORP7. Cell Tissue Res 315, 39–57 (2004). https://doi.org/10.1007/s00441-003-0817-y

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