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The NKG2D ligands RAE-1δ and RAE-1ε differ with respect to their receptor affinity, expression profiles and transcriptional regulation.

Cédile O, Popa N, Pollet-Villard F, Garmy N, Ibrahim EC, Boucraut J - PLoS ONE (2010)

Bottom Line: The interest of this result is especially related to the observation that RAE-1δ is a weak NKG2D ligand compared to RAE-1ε.Since the protein expression correlated with the level of transcripts for each rae-1 gene, we postulated that transcriptional regulation is one of the main processes explaining the difference between RAE-1δ and RAE-1ε expression.Immune function would be mainly exerted by RAE-1ε and non-immune function by RAE-1δ.

View Article: PubMed Central - PubMed

Affiliation: CRN2M, CNRS UMR 6231, Université de la Méditerranée, Université Paul Cézanne, Faculté de Médecine, Marseille, France.

ABSTRACT

Background: RAE-1 is a ligand of the activating receptor NKG2D expressed by NK cells, NKT, γδT and some CD8(+)T lymphocytes. RAE-1 is overexpressed in tumor cell lines and its expression is induced after viral infection and genotoxic stress. We have recently demonstrated that RAE-1 is expressed in the adult subventricular zone (SVZ) from C57BL/6 mice. RAE-1 is also expressed in vitro by neural stem/progenitor cells (NSPCs) and plays a non-immune role in cell proliferation. The C57BL/6 mouse genome contains two rae-1 genes, rae-1δ and rae-1ε encoding two different proteins. The goals of this study are first to characterize the in vivo and in vitro expression of each gene and secondly to elucidate the mechanisms underlying their respective expression, which are far from known.

Principal findings: We observed that Rae-1δ and Rae-1ε transcripts are differentially expressed according to tissues, pathological conditions and cell lines. Embryonic tissue and the adult SVZ mainly expressed Rae-1δ transcripts. The NSPCs derived from the SVZ also mainly expressed RAE-1δ. The interest of this result is especially related to the observation that RAE-1δ is a weak NKG2D ligand compared to RAE-1ε. On the contrary, cell lines expressed either similar levels of RAE-1δ and RAE-1ε proteins or only RAE-1ε. Since the protein expression correlated with the level of transcripts for each rae-1 gene, we postulated that transcriptional regulation is one of the main processes explaining the difference between RAE-1δ and RAE-1ε expression. We indeed identified two different promoter regions for each gene: one mainly involved in the control of rae-1δ gene expression and the other in the control of rae-1ε expression.

Conclusions/significance: RAE-1δ and RAE-1ε differ with respect to their function and the control of their expression. Immune function would be mainly exerted by RAE-1ε and non-immune function by RAE-1δ.

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Related in: MedlinePlus

Quantification of all Rae-1 transcripts in vivo and in vitro.A, Schematic representation of rae-1 genes with exons (boxes), transcription start sites (bold arrows), intron splicing (bold doted lines) and alternative splicing (thin doted lines). Prediction of all Rae-1δ and Rae-1ε transcripts under the control of either promoter 1 or promoter 2 and position of sets of primers allowing the detection and quantification of each transcript. B, RT-qPCR of each transcript in 10-days-old embryo (E10), subventricular zone (SVZ), liver, olfactory bulbs (OB) from control or axotomized mice, lumbar spinal cord (LSC) from mice suffering EAE and neurospheres (NS), C57sv and NOE cell lines. Results are expressed relatively to GAPDH as endogenous control.
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pone-0013466-g006: Quantification of all Rae-1 transcripts in vivo and in vitro.A, Schematic representation of rae-1 genes with exons (boxes), transcription start sites (bold arrows), intron splicing (bold doted lines) and alternative splicing (thin doted lines). Prediction of all Rae-1δ and Rae-1ε transcripts under the control of either promoter 1 or promoter 2 and position of sets of primers allowing the detection and quantification of each transcript. B, RT-qPCR of each transcript in 10-days-old embryo (E10), subventricular zone (SVZ), liver, olfactory bulbs (OB) from control or axotomized mice, lumbar spinal cord (LSC) from mice suffering EAE and neurospheres (NS), C57sv and NOE cell lines. Results are expressed relatively to GAPDH as endogenous control.

Mentions: A, RT-PCR analysis of Rae-1δ transcripts using an exon 1-exon 6δ primer pair or an exon 5-exon 6δ primer pair and of Rae-1ε transcripts using an exon 1-exon 6ε primer pair or an exon 5-exon 6ε primer pair (see schematic in Fig. 6). We compared the PCR products obtained from cell lines (C57sv, ES, NOE) and healthy tissues (10-days-old embryo (E10), liver, heart (He) and brain (Br)); NTC, non template control.


The NKG2D ligands RAE-1δ and RAE-1ε differ with respect to their receptor affinity, expression profiles and transcriptional regulation.

Cédile O, Popa N, Pollet-Villard F, Garmy N, Ibrahim EC, Boucraut J - PLoS ONE (2010)

Quantification of all Rae-1 transcripts in vivo and in vitro.A, Schematic representation of rae-1 genes with exons (boxes), transcription start sites (bold arrows), intron splicing (bold doted lines) and alternative splicing (thin doted lines). Prediction of all Rae-1δ and Rae-1ε transcripts under the control of either promoter 1 or promoter 2 and position of sets of primers allowing the detection and quantification of each transcript. B, RT-qPCR of each transcript in 10-days-old embryo (E10), subventricular zone (SVZ), liver, olfactory bulbs (OB) from control or axotomized mice, lumbar spinal cord (LSC) from mice suffering EAE and neurospheres (NS), C57sv and NOE cell lines. Results are expressed relatively to GAPDH as endogenous control.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC2957426&req=5

pone-0013466-g006: Quantification of all Rae-1 transcripts in vivo and in vitro.A, Schematic representation of rae-1 genes with exons (boxes), transcription start sites (bold arrows), intron splicing (bold doted lines) and alternative splicing (thin doted lines). Prediction of all Rae-1δ and Rae-1ε transcripts under the control of either promoter 1 or promoter 2 and position of sets of primers allowing the detection and quantification of each transcript. B, RT-qPCR of each transcript in 10-days-old embryo (E10), subventricular zone (SVZ), liver, olfactory bulbs (OB) from control or axotomized mice, lumbar spinal cord (LSC) from mice suffering EAE and neurospheres (NS), C57sv and NOE cell lines. Results are expressed relatively to GAPDH as endogenous control.
Mentions: A, RT-PCR analysis of Rae-1δ transcripts using an exon 1-exon 6δ primer pair or an exon 5-exon 6δ primer pair and of Rae-1ε transcripts using an exon 1-exon 6ε primer pair or an exon 5-exon 6ε primer pair (see schematic in Fig. 6). We compared the PCR products obtained from cell lines (C57sv, ES, NOE) and healthy tissues (10-days-old embryo (E10), liver, heart (He) and brain (Br)); NTC, non template control.

Bottom Line: The interest of this result is especially related to the observation that RAE-1δ is a weak NKG2D ligand compared to RAE-1ε.Since the protein expression correlated with the level of transcripts for each rae-1 gene, we postulated that transcriptional regulation is one of the main processes explaining the difference between RAE-1δ and RAE-1ε expression.Immune function would be mainly exerted by RAE-1ε and non-immune function by RAE-1δ.

View Article: PubMed Central - PubMed

Affiliation: CRN2M, CNRS UMR 6231, Université de la Méditerranée, Université Paul Cézanne, Faculté de Médecine, Marseille, France.

ABSTRACT

Background: RAE-1 is a ligand of the activating receptor NKG2D expressed by NK cells, NKT, γδT and some CD8(+)T lymphocytes. RAE-1 is overexpressed in tumor cell lines and its expression is induced after viral infection and genotoxic stress. We have recently demonstrated that RAE-1 is expressed in the adult subventricular zone (SVZ) from C57BL/6 mice. RAE-1 is also expressed in vitro by neural stem/progenitor cells (NSPCs) and plays a non-immune role in cell proliferation. The C57BL/6 mouse genome contains two rae-1 genes, rae-1δ and rae-1ε encoding two different proteins. The goals of this study are first to characterize the in vivo and in vitro expression of each gene and secondly to elucidate the mechanisms underlying their respective expression, which are far from known.

Principal findings: We observed that Rae-1δ and Rae-1ε transcripts are differentially expressed according to tissues, pathological conditions and cell lines. Embryonic tissue and the adult SVZ mainly expressed Rae-1δ transcripts. The NSPCs derived from the SVZ also mainly expressed RAE-1δ. The interest of this result is especially related to the observation that RAE-1δ is a weak NKG2D ligand compared to RAE-1ε. On the contrary, cell lines expressed either similar levels of RAE-1δ and RAE-1ε proteins or only RAE-1ε. Since the protein expression correlated with the level of transcripts for each rae-1 gene, we postulated that transcriptional regulation is one of the main processes explaining the difference between RAE-1δ and RAE-1ε expression. We indeed identified two different promoter regions for each gene: one mainly involved in the control of rae-1δ gene expression and the other in the control of rae-1ε expression.

Conclusions/significance: RAE-1δ and RAE-1ε differ with respect to their function and the control of their expression. Immune function would be mainly exerted by RAE-1ε and non-immune function by RAE-1δ.

Show MeSH
Related in: MedlinePlus