scholarly journals A mutated retinoic acid receptor-alpha exhibiting dominant-negative activity alters the lineage development of a multipotent hematopoietic cell line.

1992 ◽  
Vol 6 (12a) ◽  
pp. 2258-2269 ◽  
Author(s):  
S Tsai ◽  
S Bartelmez ◽  
R Heyman ◽  
K Damm ◽  
R Evans ◽  
...  
Keyword(s):  
Retinoic Acid ◽  
Cell Line ◽  
Retinoic Acid Receptor ◽  
Dominant Negative ◽  
Hematopoietic Cell ◽  
Retinoic Acid Receptor Alpha ◽  
Acid Receptor ◽  
Receptor Alpha ◽  
Hematopoietic Cell Line ◽  
Dominant Negative Activity

Retinoic acid-resistant HL-60 cells exclusively contain mutant retinoic acid receptor-alpha

Blood ◽  
1994 ◽  
Vol 83 (11) ◽  
pp. 3298-3302 ◽  
Author(s):  
YP Li ◽  
F Said ◽  
RE Gallagher
Keyword(s):  
Retinoic Acid ◽  
Retinoic Acid Receptor ◽  
Hot Spot ◽  
Dominant Negative ◽  
Reaction Products ◽  
Retinoic Acid Receptor Alpha ◽  
Restriction Endonuclease Site ◽  
Acid Receptor ◽  
Receptor Alpha ◽  
Alpha Gene

Abstract Sequence analysis of the retinoic acid receptor-alpha (RAR alpha) gene from a subline of HL-60 cells (RA-res) stably resistant to all-trans retinoic acid (RA) disclosed a single-base change in codon number 411, the same C to T transition previously reported in an independently selected HL-60 RA resistant clone by Robertson et al (Blood 80:1885, 1992). This mutation eliminates a FokI restriction endonuclease site. Using primers framing this mutation in exon 9 of the RAR alpha gene, we showed that polymerase chain reaction products amplified from either mRNA or genomic DNA templates from the RA-res subline were completely resistant to FokI digestion whereas those from wild-type (wt) HL-60 cells could be digested to completion. The lack of a normal allele in the RA-res cells was confirmed by mixing experiments and hybridization analyses. Southern blot analysis of DNA from the RA-res and wt cells versus control placental DNA indicated that the RAR alpha gene is not haploid. The independent isolation of the same RAR alpha mutation in different laboratories suggests either that the mutation exits in a small subpopulation in the wt line or that this is a mutational “hot spot.” Furthermore, the results indicate that if a dominant negative mode of resistance is involved in the RA-res subline, this must involve interference with the function of heterologous receptor proteins such as the retinoid X receptors. The lack of any normal RAR alpha in this subline may facilitate studies of the mode of action of retinoids.

scholarly journals A58 REGULATION OF INTESTINAL EPITHELIAL THYMIC STROMAL LYMPHOPOIETIN GENE EXPRESSION BY RETINOIC ACID RECEPTOR ALPHA

2020 ◽  
Vol 3 (Supplement_1) ◽  
pp. 68-69
Author(s):  
R Mahmood ◽  
R Chan ◽  
P Beck ◽  
H Jijon
Keyword(s):  
Retinoic Acid ◽  
Cell Line ◽  
Retinoic Acid Receptor ◽  
Thymic Stromal Lymphopoietin ◽  
Immune Homeostasis ◽  
Intestinal Epithelial ◽  
Retinoic Acid Receptor Alpha ◽  
Acid Receptor ◽  
Receptor Alpha ◽  
Selective Agonist

Abstract Background We have targeted the retinoic acid receptor alpha (RARα) isoform in murine intestinal epithelial cells (IECs) to explore the role of epithelial intrinsic retinoid signaling in intestinal immune homeostasis. We noted a significant decrease in the CD11c+ and CD103+ dendritic cell (DC) subpopulations in the intestines of intestinal-epithelial specific RARα-deficient (RARα villin) mice. In the present study, we identified Thymic Stromal Lymphopoietin (TSLP) as a contributing factor that influences intrinsic RARα signaling within IECs. This cytokine preferentially stimulates CD103+ DCs to a more tolerogenic phenotype in the GI tract, induces Tregs and drives TH2 polarization. TSLP also influences immune homeostasis by modulating the activation of myeloid cells, inhibition of proinflamamtory DCs, and production of IL-12. Notably, TSLP levels are decreased in Crohn’s disease (CD) where inflammation is driven by IL-12. Aims We hypothesize that RA intrinsic signaling within IECs modulates TSLP expression via activation of RARα. We sought to generate a RARα -/- murine intestinal epithelial cell line utilizing the CRISPR/CAS9 system to examine the effects of RARa ablation. Second, we aim to establish RARα’s role in regulating intestinal epithelial TSLP expression in both the RARα villin mice and the CRISPR-generated RARα -/- cell line. Methods Exon 5 in the open reading frame of the RARα gene in Mode-K murine IECs was targeted using CRISPR/CAS9. CRISPR cleavage and knockdown of RARα activity was confirmed via surveyor/luciferase assays. The cell lines were phenotyped with regards to their morphology, proliferative ability, and viability. Next, we examined TSLP mRNA expression in the colons of RARα villin mice and the CRISPR RARα -/- knockout cell line via qRT-PCR under baseline and stimulated conditions. Wild-type (WT) and RARα-/- cells were stimulated with RARα-selective agonist, BMS753, and muramyl dipeptide (MDP), a NOD2-selective agonist. Results TSLP expression in surface colonic epithelium was elevated 2.3-fold in RARα villin mice compared to WT litter mates. TSLP expression was elevated 10-fold in the Mode-K RARα-/- cells versus parent cells at baseline. BMS753 stimulation resulted in a 15-fold increase in TSLP expression in the RARα-/- cells compared to baseline. When stimulated with MDP, a NOD2-selective agonist known to stimulate TSLP expression, TSLP expression was elevated 60-fold in the RARα-/- cells, but significantly impaired compared to the WT cells (400-fold). Conclusions TSLP expression is controlled by RARα in colonic IECs where it may act as a repressor of TSLP promoter transactivation under baseline conditions. This suggests an important role for RA on myeloid and T cell function via effects on intestinal epithelial TSLP expression. Funding Agencies CCCUniversity of Calgary, Department of Medicine

scholarly journals Retinoic acid-resistant HL-60 cells exclusively contain mutant retinoic acid receptor-alpha

Blood ◽  
1994 ◽  
Vol 83 (11) ◽  
pp. 3298-3302 ◽  
Author(s):  
YP Li ◽  
F Said ◽  
RE Gallagher
Keyword(s):  
Retinoic Acid ◽  
Retinoic Acid Receptor ◽  
Hot Spot ◽  
Dominant Negative ◽  
Reaction Products ◽  
Retinoic Acid Receptor Alpha ◽  
Restriction Endonuclease Site ◽  
Acid Receptor ◽  
Receptor Alpha ◽  
Alpha Gene

Sequence analysis of the retinoic acid receptor-alpha (RAR alpha) gene from a subline of HL-60 cells (RA-res) stably resistant to all-trans retinoic acid (RA) disclosed a single-base change in codon number 411, the same C to T transition previously reported in an independently selected HL-60 RA resistant clone by Robertson et al (Blood 80:1885, 1992). This mutation eliminates a FokI restriction endonuclease site. Using primers framing this mutation in exon 9 of the RAR alpha gene, we showed that polymerase chain reaction products amplified from either mRNA or genomic DNA templates from the RA-res subline were completely resistant to FokI digestion whereas those from wild-type (wt) HL-60 cells could be digested to completion. The lack of a normal allele in the RA-res cells was confirmed by mixing experiments and hybridization analyses. Southern blot analysis of DNA from the RA-res and wt cells versus control placental DNA indicated that the RAR alpha gene is not haploid. The independent isolation of the same RAR alpha mutation in different laboratories suggests either that the mutation exits in a small subpopulation in the wt line or that this is a mutational “hot spot.” Furthermore, the results indicate that if a dominant negative mode of resistance is involved in the RA-res subline, this must involve interference with the function of heterologous receptor proteins such as the retinoid X receptors. The lack of any normal RAR alpha in this subline may facilitate studies of the mode of action of retinoids.

Retinoic Acid Receptor Alpha Amplifications and Retinoic Acid Sensitivity in Breast Cancers

2013 ◽  
Vol 13 (5) ◽  
pp. 401-408 ◽  
Author(s):  
Samar Alsafadi ◽  
Caroline Even ◽  
Coralie Falet ◽  
Aicha Goubar ◽  
Frédéric Commo ◽  
...  
Keyword(s):  
Retinoic Acid ◽  
Retinoic Acid Receptor ◽  
Breast Cancers ◽  
Retinoic Acid Receptor Alpha ◽  
Acid Sensitivity ◽  
Acid Receptor ◽  
Receptor Alpha

Retinoic Acid Receptor Alpha (RARα) Fusion Genes in Leukemia

2017 ◽  
pp. 271-285
Author(s):  
Ganesan Padmavathi ◽  
Javadi Monisha ◽  
Anand Anip ◽  
Krishan Kumar Thakur ◽  
Ajaikumar B. Kunnumakkara
Keyword(s):  
Retinoic Acid ◽  
Retinoic Acid Receptor ◽  
Fusion Genes ◽  
Retinoic Acid Receptor Alpha ◽  
Acid Receptor ◽  
Receptor Alpha

Inhibitory Effects of Retinoic Acid Receptor Alpha Stimulants on Murine Cataractogenesis through Suppression of Deregulated Calpains

2007 ◽  
Vol 48 (5) ◽  
pp. 2224 ◽  
Author(s):  
Nami Nishikiori ◽  
Makoto Osanai ◽  
Hideki Chiba ◽  
Takashi Kojima ◽  
Hiroshi Ohguro ◽  
...  
Keyword(s):  
Retinoic Acid ◽  
Retinoic Acid Receptor ◽  
Inhibitory Effects ◽  
Retinoic Acid Receptor Alpha ◽  
Acid Receptor ◽  
Receptor Alpha

scholarly journals Characterization of a functional promoter for the human retinoic acid receptor-alpha (hRAR-α)

1990 ◽  
Vol 18 (23) ◽  
pp. 6799-6806 ◽  
Author(s):  
Nigel J. Brand ◽  
Martin Petkovich ◽  
Pierre Chambon
Keyword(s):  
Retinoic Acid ◽  
Retinoic Acid Receptor ◽  
Retinoic Acid Receptor Alpha ◽  
Acid Receptor ◽  
Receptor Alpha
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