Rod photoreceptor-specific gene expression in human retinoblastoma cells.
Retinoblastoma cells in culture have previously been shown to express cone-specific genes but not their rod counterparts. We have detected the messages for the rod alpha, beta, and gamma subunits of cGMP phosphodiesterase (PDE), the rod alpha subunit of transducin, rod opsin, and the cone alpha' subunit of PDE in RNA of human Y-79 retinoblastoma cells by reverse transcription-PCR. Quantitative analysis of the mRNAs for the rod alpha and cone alpha' PDE subunits revealed that they were expressed at comparable levels; however, the transcript encoding the rod beta PDE subunit was 10 times more abundant in these cells. Northern hybridization analysis of Y-79 cell RNA confirmed the presence of the transcripts for rod and cone PDE catalytic subunits. To test whether the transcriptional machinery required for the expression of rod-specific genes was endogenous in Y-79 retinoblastoma cells, cultures were transfected with a construct containing the promoter region of the rod beta PDE subunit gene attached to the firefly luciferase reporter vector. Significant levels of reporter enzyme activity were observed in the cell lysates. Our results demonstrate that the Y-79 retinoblastoma cell line is a good model system for the study of transcriptional regulation of rod-specific genes.
Di Polo
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Department of Ophthalmology, University of California, Los Angeles School of Medicine 90024, USA.
Farber
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DB
D015106:3',5'-Cyclic-GMP Phosphodiesterases / Q000096:biosynthesis / Q000235:genetics; D001483:Base Sequence; D002460:Cell Line; D003001:Cloning, Molecular; D017931:DNA Primers; D005134:Eye Neoplasms / Q000235:genetics / Q000378:metabolism*; D015870:Gene Expression; D006801:Humans; D046911:Macromolecular Substances; D008969:Molecular Sequence Data; D016376:Oligonucleotides, Antisense; D016133:Polymerase Chain Reaction; D012333:RNA, Messenger / Q000096:biosynthesis / Q000302:isolation & purification; D012334:RNA, Neoplasm / Q000302:isolation & purification; D015183:Restriction Mapping; D017948:Retinal Rod Photoreceptor Cells / Q000378:metabolism*; D012175:Retinoblastoma / Q000235:genetics / Q000378:metabolism*; D012243:Rhodopsin / Q000235:genetics; D014162:Transfection; D014407:Tumor Cells, Cultured
D017931:DNA Primers; D046911:Macromolecular Substances; D016376:Oligonucleotides, Antisense; D012333:RNA, Messenger; D012334:RNA, Neoplasm; D012243:Rhodopsin; D015106:3',5'-Cyclic-GMP Phosphodiesterases