A 350 bp region of the proximal promoter of Rds drives cell-type specific gene expression.
RDS (retinal degeneration slow) is a photoreceptor-specific tetraspanin protein required for the biogenesis and maintenance of rod and cone outer segments. Mutations in the Rds gene are associated with multiple forms of rod- and cone-dominant retinal degeneration. To gain more insight into the mechanisms underlying the regulation of this gene, the identification of regulatory sequences within the promoter of Rds was undertaken. A 3.5 kb fragment of the 5' flanking region of the mouse Rds gene was isolated and binding sites for Crx, Otx2, Nr2e3, RXR family members, Mef2C, Esrrb, NF1, AP1, and SP1 in addition to several E-boxes, GC-boxes and GAGA-boxes were identified. Crx binding sequences were conserved in all mammalian species examined. Truncation expression analysis of the Rds promoter region in Y-79 retinoblastoma cells showed maximal activity in the 350 bp proximal promoter region. We also show that inclusion of more distal fragments reduced promoter activity to the basal level, and that the promoter activities are cell-type and direction specific. Co-transfection with Nrl increased promoter activity, suggesting that this gene positively regulates Rds expression. Based on these findings, a relatively small fragment of the Rds promoter may be useful in future gene transfer studies to drive gene expression in photoreceptors.
Cai
Xue
X
Department of Cell Biology, University Oklahoma Health Sciences Center, 940 Stanton L. Young Blvd, BMSB 781, Oklahoma City, OK 73104, USA.
Conley
Shannon M
SM
Cheng
Tong
T
Al-Ubaidi
Muayyad R
MR
Naash
Muna I
MI
D024506:5' Flanking Region / Q000235:genetics; D000818:Animals; D001665:Binding Sites; D015153:Blotting, Western; D003001:Cloning, Molecular; D019084:Fluorescent Antibody Technique, Indirect; D005786:Gene Expression Regulation / Q000502:physiology*; D007381:Intermediate Filament Proteins / Q000235:genetics*; D008562:Membrane Glycoproteins / Q000235:genetics*; D051379:Mice; D008807:Mice, Inbred BALB C; D008810:Mice, Inbred C57BL; D009419:Nerve Tissue Proteins / Q000235:genetics*; D064531:Peripherins; D020419:Photoreceptor Cells, Vertebrate / Q000378:metabolism; D010957:Plasmids; D011401:Promoter Regions, Genetic / Q000235:genetics*; D038621:Regulatory Sequences, Ribonucleic Acid / Q000235:genetics; D012162:Retinal Degeneration / Q000235:genetics*; D019572:Retinal Neoplasms / Q000235:genetics; D012175:Retinoblastoma / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014157:Transcription Factors / Q000235:genetics; D014162:Transfection; D014407:Tumor Cells, Cultured
D007381:Intermediate Filament Proteins; D008562:Membrane Glycoproteins; D009419:Nerve Tissue Proteins; D064531:Peripherins; C578996:Prph2 protein, mouse; D038621:Regulatory Sequences, Ribonucleic Acid; D014157:Transcription Factors
Mol Vis. 2006 Apr 07;12:298-309
16617297
Proc Natl Acad Sci U S A. 1995 Apr 25;92(9):4016-20
7732024
Proc Natl Acad Sci U S A. 1993 Jun 15;90(12):5499-503
8516292
Anal Biochem. 2005 Nov 15;346(2):289-94
16213455
Exp Eye Res. 2009 Sep;89(3):365-72
19379737
Gene Ther. 2007 Feb;14(4):292-303
17024105
J Virol. 2007 Oct;81(20):11372-80
17699581
Nat Genet. 2001 May;28(1):92-5
11326284
PLoS One. 2009;4(4):e5290
19390689
Invest Ophthalmol Vis Sci. 1993 Apr;34(5):1659-76
8473105
Curr Biol. 2000 Mar 23;10(6):301-10
10744971
Hum Mol Genet. 2004 Aug 1;13(15):1563-75
15190009
Nucleic Acids Res. 1987 Oct 26;15(20):8125-48
3313277
J Biol Chem. 2004 May 7;279(19):19800-7
15001570
Invest Ophthalmol Vis Sci. 2004 Aug;45(8):2514-21
15277471
Mol Cell Biol. 2007 Dec;27(23):8318-29
17908793
Nucleic Acids Res. 1999 Oct 1;27(19):3792-8
10481017
Cell. 1997 Nov 14;91(4):531-41
9390562
J Biol Chem. 2001 Apr 27;276(17):14037-43
11278688
J Biol Chem. 1991 May 25;266(15):9667-72
1827795
Invest Ophthalmol Vis Sci. 2007 Sep;48(9):3954-61
17724172
J Cell Biol. 2006 Apr 10;173(1):59-68
16585269
J Biol Chem. 2004 Nov 19;279(47):49010-8
15347652
Hum Gene Ther. 2008 Oct;19(10):979-90
18774912
Invest Ophthalmol Vis Sci. 2004 Mar;45(3):764-8
14985288
Nat Genet. 2000 Jul;25(3):306-10
10888879
Proc Natl Acad Sci U S A. 2008 Sep 30;105(39):15112-7
18809924
J Mol Neurosci. 1998 Jun;10(3):235-50
9770645
J Biol Chem. 2001 Sep 28;276(39):36557-65
11333267
Vis Neurosci. 2006 Nov-Dec;23(6):917-29
17266784
Exp Eye Res. 1993 Apr;56(4):497-507
8500562
Hum Mutat. 1996;8(4):297-303
8956033
FASEB J. 2010 Apr;24(4):1178-91
19952284
Proc Natl Acad Sci U S A. 2006 Apr 18;103(16):6218-23
16606843
Hum Mol Genet. 2006 Sep 1;15(17):2588-602
16868010
Biochem Biophys Res Commun. 1998 Sep 8;250(1):175-80
9735352
Gene. 2002 Oct 2;298(2):173-82
12426105
Vis Neurosci. 2005 Sep-Oct;22(5):575-86
16332268
Proc Natl Acad Sci U S A. 2007 Jan 30;104(5):1679-84
17242361
Neuron. 1997 Nov;19(5):1017-30
9390516
Invest Ophthalmol Vis Sci. 2003 Sep;44(9):4076-85
12939331
Mamm Genome. 2008 Sep;19(9):623-33
18763016
J Biol Chem. 1998 Mar 6;273(10):5591-8
9488687
Mol Ther. 2003 Jun;7(6):774-81
12788651
J Gene Med. 2009 May;11(5):444-53
19263463
J Neurochem. 2006 Apr;97(2):556-66
16539656
Proc Natl Acad Sci U S A. 1996 Jan 9;93(1):191-5
8552602
Gene. 2004 Sep 15;339:139-47
15363854
N Engl J Med. 2008 May 22;358(21):2231-9
18441371
J Mol Biol. 1987 Jul 20;196(2):261-82
3656447
Hum Gene Ther. 2003 Nov 20;14(17):1663-71
14633408
PLoS One. 2007 Jul 25;2(7):e643
17653270
Brain Res. 2008 Feb 4;1192:114-33
17662965
Mol Ther. 2005 Dec;12(6):1072-82
16226919
Hum Mol Genet. 2005 Mar 15;14(6):747-64
15689355
Methods Enzymol. 1987;152:704-20
2889129
Hum Mol Genet. 2004 Jul 15;13(14):1487-503
15163632
Hum Gene Ther. 2006 Aug;17(8):845-58
16942444
Mol Vis. 1997 Dec 22;3:15
9479006
J Neurochem. 1996 Dec;67(6):2494-504
8931483