title: Identification of RPE65 in transformed kidney cells.
abstract: 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.
authors.affiliation: Department of Ophthalmology, Medical University of South Carolina, Charleston 29425, USA. majx@musc.edu
authors.forename: J X
authors.initials: JX
authors.lastname: Ma
authors.forename: D
authors.initials: D
authors.lastname: Zhang
authors.forename: M
authors.initials: M
authors.lastname: Laser
authors.forename: N A
authors.initials: NA
authors.lastname: Brownlee
authors.forename: G G
authors.initials: GG
authors.lastname: Re
authors.forename: D J
authors.initials: DJ
authors.lastname: Hazen-Martin
authors.forename: T M
authors.initials: TM
authors.lastname: Redmond
authors.forename: R K
authors.initials: RK
authors.lastname: Crouch
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal; D001483:Base Sequence; D019556:COS Cells; D002352:Carrier Proteins; D002417:Cattle; D002461:Cell Line, Transformed; D002522:Chlorocebus aethiops; D018076:DNA, Complementary; D005136:Eye Proteins; D006801:Humans; D007668:Kidney; D008969:Molecular Sequence Data; D014176:Protein Biosynthesis; D011506:Proteins / Q000032:analysis / Q000235:genetics*; D012333:RNA, Messenger / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction; D016415:Sequence Alignment; D017386:Sequence Homology, Amino Acid; D014158:Transcription, Genetic; D014562:Caudata; D019745:cis-trans-Isomerases; D000099178:Retinoid Isomerohydrolase
chemical_list: D000911:Antibodies, Monoclonal; D002352:Carrier Proteins; D018076:DNA, Complementary; D005136:Eye Proteins; D011506:Proteins; D012333:RNA, Messenger; D019745:cis-trans-Isomerases; D000099178:Retinoid Isomerohydrolase