Identification of RPE65 in transformed kidney cells1

FEBS Letters

Brownlee, Noel A.²  Re, Gian G.²  Redmond, T.Michael³  Crouch, Rosalie K.¹  Hazen-Martin, Debra J.²  Laser, Martin¹  Zhang, Dongchang¹  Ma, Jian-xing¹ 

[1] Department of Ophthalmology, 7th. Floor, Medical University of South Carolina, 167 Ashley Ave., Charleston, SC 29425, USA;Department of Pathology and Laboratory Medicine, Medical University of South Carolina, 167 Ashley Ave., Charleston, SC 29425, USA;Laboratory of Retinal Cell and Molecular Biology, National Eye Institute, NIH, 9000 Rockville Pike, Bethesda, MD 20892, USA

关键词: COS cell;    Kidney;    Retinal pigment epithelium;    Retinoid;    RPE65;    Tumor;    arCSRD;    autosomal recessive childhood onset severe retinal dystrophy;    GAPDH;    glyceraldehyde-3-phosphate dehydrogenase;    LCA;    Leber's congenital amaurosis;    RBP;    retinol-binding protein;    RPE;    retinal pigment epithelium;    RT-PCR;    reverse transcription and polymerase chain reaction;   

DOI  :  10.1016/S0014-5793(99)00606-7

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

The protein RPE65 has an important role in retinoid processing and/or retinoid transport in the eye. Retinoids are involved in cell differentiation, embryogenesis and carcinogenesis. Since the kidney is known as an important site for retinoid metabolism, the expression of RPE65 in normal kidney and transformed kidney cells has been examined. The RPE65 mRNA was detected in transformed kidney cell lines including the human embryonic kidney cell line HEK293 and the African green monkey kidney cell lines COS-1 and COS-7 by reverse transcription PCR. In contrast, it was not detected in human primary kidney cells or monkey kidney tissues under the same PCR conditions. The RPE65 protein was also identified in COS-7 and HEK293 cells by Western blot analysis using a monoclonal antibody to RPE65, but not in the primary kidney cells or kidney tissues. The RPE65 cDNA containing the full-length encoding region was amplified from HEK293 and COS-7 cells. DNA sequencing showed that the RPE65 cDNA from HEK293 cells is identical to the RPE65 cDNA from the human retinal pigment epithelium. The RPE65 from COS-7 cells shares 98 and 99% sequence identity with human RPE65 at the nucleotide and amino acid levels, respectively. Moreover, the RPE65 mRNA was detected in three out of four renal tumor cultures analyzed including congenital mesoblastic nephroma and clear cell sarcoma of the kidney. These results demonstrated that transformed kidney cells express this retinoid processing protein, suggesting that these transformed cells may have an alternative retinoid metabolism not present in normal kidney cells.

【 授权许可】

Unknown