Nrl 18185

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Gene ID: 18185
Gene symbol: nrl
Gene Description: neural retina leucine zipper gene
Aliases: AW492574; D14H14S46E
Human Ortholog Gene ID: [4901]
Reviewer Judgement: TF Gene; TF Gene Candidate
Functional Classification: DNA-Binding: non-sequence-specific;

Transcription Factor Binding: tf co-factor binding; DNA-Binding: sequence-specific

DBD Protein Group Classification: Zipper-Type Group
DBD Protein Group/Family Classification: Zipper-Type Group / Leucine Zipper Family
Homolog Cluster(s): 16658

Evidence:

Pubmed ID Function Species Evidence Strength Reviewer's Comments Reviewer
[8552602] DNA Binding Human Medium EMSA shows two shifted bands for rhodopsin NRE fragment RhoP, not competitively inhibited by a slight mutated mRhoP, bound by Nrl. Presence of RhoP bound to Nrl confirmed by antibody. Human NRL was used in this study. Warren Cheung
[10887186] TF PPI Mouse Weak Yeast two-hybrid shows NRL leucine zipper domain interacting with CRX. 35S CRX binds to GST-NRL but not GST, in vitro SDS-PAGE. CRX domains that interact strongly with NRL were found via CRX deletion constructs and pull-down assays. Leucine Zipper domain of NRL needed to bind CRX. Warren Cheung
[15001570] Transactivation Mouse Medium Nrl activates PDE6A-promoter luc reporter. Also shows synergistic activation with Crx which is abolished by DD10. Quantitative PCR shows Nrl required for Pde6a transcription. Warren Cheung
[12566383] Transactivation Human; Mouse Strong Nrl transactivates the rhodopsin promoter reporter construct p130-Luc in transient transfections of CV-1 cells. Magdalena Swanson
[15001570] Transactivation Human Strong Nrl increased PDE6A promoter activity (plasmid pGL3-300) in transient transfection experiments in HEK 293 cells. Magdalena Swanson
[17335001] DNA Binding Human Strong In competition EMSA, NRL bound to the rhodopsin-NRE sequence but not to a non-specific oligonucleotide. Magdalena Swanson
[8552602] Co-activation; DNA Binding; Transactivation Human; Other Medium GeneID: 18185

Referring to PMID: 8552602

Nrl may be a transcription factor, which binds the NRE interaction element upstream of the human rhodopsin gene. In EMSA assays, a 24-bp oligonucleotide containing the rhodopsin NRE, exhibits shifts in mobility upon incubation with bovine retinal nuclear extract or Y79 human retinoblastoma cell extract. A supershift is also observed when antibody for Nrl is used. The interaction is also competed off with the addition of unlabeled human rhodopsin. No significant competition is observed when unlabeled NRE is used as a competitive inhibitor, even when used in 500-fold molar excess.

Nrl transactivates the Rhodopsin Promoter. Nrl expression vector is cotransfected into CV-1 cells with luciferase constructs containing rhodopsin promoter fragments. Cotransfection of pMT-NRL with the p84- Luc construct, which contains a complete NRE, consistently leads to 3-fold stimulation of luciferase activity. p61-Luc, which does not contain a complete NRE, shows no induction upon Nrl expression. p130-Luc and p176-Luc also show responsiveness to Nrl, whereas p225-Luc and p4907-Luc are unresponsive to Nrl-mediated transactivation.

Stuart Lithwick
[8552602] DNA Binding Human; Other Medium Electrophoretic mobility-shift analysis reveals that a protein(s) in nuclear extracts from bovine retina and the Y79 human retinoblastoma cell line binds to a conserved Nrl response element (NRE) in the upstream promoter region of the rhodopsin gene. Nrl or an antigenically similar protein is shown to be part of the bound protein complex by supershift experiments using Nrl-specific antiserum. Coexpression of Nrl is shown to transactivate the rhodopsin promoter. Amy Ticoll
[8939891] DNA Binding Other Strong To investigate the role of Nrl in rhodopsin regulation, E8 and E17 retinal cell cultures are cotransfected with bRho constructs with an expression vector containing the human NRL cDNA (pMT-NRL). Cotransfection of pMT-NRL results in induction of reporter gene activity with constructs that contained an intact NRE. Constructs that lack the NRE or contained a mutated NRE have little induction, in both E8 and E17 cells. Using a yeast one-hybrid system, Nrl is shown to bind to the NRE in vivo and regulate expression of a reporter gene. Amy Ticoll
Additional Reviewer Comments:
Reviewer: Amy Ticoll

Using the yeast one-hybrid system, Nrl can bind to the NRE in vivo and regulate a reporter gene. EMSA and coexpression reveal that Nrl plays a role in binding and activating the Nrl element. Binding specificity was demonstrated with cotransfection of pMT-NRL.

Reviewer: Magdalena Swanson

Nrl is a transcription factor. It activates transcription from reporter constructs in transient transfection experiments of HEK 293 and CV-1 cells. It binds to specific DNA sequences, as demonstrated by a competition EMSA.

Reviewer: Stuart Lithwick

GeneID: 18185 Referring to PMID: 8552602 Human Nrl is likely a transcription factor, which binds sequence-specifically to the human Rhodopsin (Rho) NRE and leads to increases in promoter activity. The human Rho NRE is bound by protein when incubated with bovine or human cell extract, as demonstrated by EMSA. The bound protein id further identified to be Nrl using a supershift assay. As well, cotransfection of human Nrl and human Rho NRE-luc expression constructs into CV-1 leads to consistent 3-fold increases in reporter activity.

Reviewer: Warren Cheung

Appears to be a TF - experiments show human Nrl binding to rhodopsin promoter, and Nrl activating PDE6A promoter. Nrl also appears to interact with CRX, with synergistic enhancement effect.


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Facts about Nrl 18185RDF feed
Gene description neural retina leucine zipper gene  +
Gene id 18,185  +
Gene judgment TF Gene  +, and TF Gene Candidate  +
Gene symbol nrl  +, AW492574  +, and D14H14S46E  +
Gene taxonomy DNA-Binding: non-sequence-specific  +, Transcription Factor Binding: tf co-factor binding  +, and DNA-Binding: sequence-specific  +
Homolog cluster 16,658  +
Protein family 3 Zipper-Type Group/21 Leucine Zipper Family  +
Protein group 3 Zipper-Type Group   +
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