PMID- 12910240
OWN - NLM
STAT- MEDLINE
DCOM- 20031016
LR  - 20131121
IS  - 1090-0535 (Electronic)
IS  - 1090-0535 (Linking)
VI  - 9
DP  - 2003 Aug 5
TI  - Down-regulation of RPE65 protein expression and promoter activity by retinoic 
      acid.
PG  - 345-54
AB  - PURPOSE: RPE65 is critical for the normal formation of 11-cis retinal and thus 
      photoreceptor function. Opsin expressed in HEK293 cells has been reported to form 
      rhodopsin on the addition of all-trans retinol, indicating that the machinery for 
      retinoid isomerization is present. RPE65 has been previously identified in HEK293 
      cells at both the RNA and protein levels. To further understand retinoid 
      metabolism in these cells and the control of RPE65 expression, HEK293 cells were 
      used as a model to determine if retinoic acid (RA) affects RPE65 promoter 
      activity. METHODS: RPE65 levels were determined by Western blots. RA regulation 
      of RPE65 promoter activity was monitored using the luciferase reporter assay 
      after transient transfection of HEK293 cells with the RPE65 promoter. Deletion 
      and truncation promoter mutants were assessed for activity. RESULTS: RA 
      down-regulates RPE65 protein expression and promoter activity. The RA receptors 
      (RARs), RARalpha, -beta, and -gamma, and the retinoid X receptors (RXRs), 
      RXRalpha, -beta, and -gamma, were all identified in these cells and shown to 
      mediate the regulation of RPE65 mRNA expression. After deletion of the AP1, AP4 
      or NF1 transcription factor binding sites, the RA down-regulation was decreased, 
      but the decrease was not associated with a single transcription factor. The 
      truncation promoter constructs P60, P153 and P257 showed increases in promoter 
      activity, indicating an inhibitory element had been removed, and the 
      down-regulatory effect of RA was decreased. CONCLUSIONS: The down-regulation of 
      RPE65 by RA is occurring at the transcription level. Multiple elements in the 
      RPE65 promoter may contribute to this regulation.
FAU - Chen, Yumei
AU  - Chen Y
AD  - Department of Ophthalmology, Medical University of South Carolina, Charleston, SC 
      29425, USA.
FAU - Ma, Jian-xing
AU  - Ma JX
FAU - Crouch, Rosalie K
AU  - Crouch RK
LA  - eng
GR  - EY04939/EY/NEI NIH HHS/United States
GR  - EY12231/EY/NEI NIH HHS/United States
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PT  - Research Support, U.S. Gov't, Non-P.H.S.
PT  - Research Support, U.S. Gov't, P.H.S.
DEP - 20030805
PL  - United States
TA  - Mol Vis
JT  - Molecular vision
JID - 9605351
RN  - 0 (Carrier Proteins)
RN  - 0 (Eye Proteins)
RN  - 0 (Proteins)
RN  - 0 (RNA, Messenger)
RN  - 0 (Receptors, Retinoic Acid)
RN  - 5688UTC01R (Tretinoin)
RN  - EC 1.13.12.- (Luciferases)
RN  - EC 3.1.1.64 (retinoid isomerohydrolase)
RN  - EC 5.2.- (cis-trans-Isomerases)
SB  - IM
MH  - Blotting, Southern
MH  - Blotting, Western
MH  - Carrier Proteins
MH  - Cell Cycle
MH  - Cell Line
MH  - Down-Regulation
MH  - Eye Proteins
MH  - Flow Cytometry
MH  - Gene Expression Regulation/*drug effects
MH  - Humans
MH  - Kidney/cytology
MH  - Luciferases/genetics/metabolism
MH  - Microscopy, Confocal
MH  - Pigment Epithelium of Eye/metabolism
MH  - Promoter Regions, Genetic/drug effects
MH  - Proteins/*genetics/metabolism
MH  - RNA, Messenger/metabolism
MH  - Receptors, Retinoic Acid/metabolism
MH  - Reverse Transcriptase Polymerase Chain Reaction
MH  - Transfection
MH  - Tretinoin/*pharmacology
MH  - cis-trans-Isomerases
EDAT- 2003/08/12 05:00
MHDA- 2003/10/17 05:00
CRDT- 2003/08/12 05:00
PHST- 2003/08/12 05:00 [pubmed]
PHST- 2003/10/17 05:00 [medline]
PHST- 2003/08/12 05:00 [entrez]
AID - v9/a48 [pii]
PST - epublish
SO  - Mol Vis. 2003 Aug 5;9:345-54.