NLS‑RARα is a novel transcriptional factor

Jiang,Kai‑Ling; Zhong,Liang; Yang,Xiao‑Qun; Ma,Peng‑Peng; Wang,Hui; Zhu,Xin‑Yu; Liu,Bei‑Zhong

doi:10.3892/ol.2017.7132

NLS‑RARα is a novel transcriptional factor

Authors:
- Kai‑Ling Jiang
- Liang Zhong
- Xiao‑Qun Yang
- Peng‑Peng Ma
- Hui Wang
- Xin‑Yu Zhu
- Bei‑Zhong Liu
View Affiliations

Affiliations: Central Laboratory of Yongchuan Hospital, Chongqing Medical University, Chongqing 402160, P.R. China, Key Laboratory of Laboratory Medical Diagnostics, Ministry of Education, Department of Laboratory Medicine, Chongqing Medical University, Chongqing 400016, P.R. China
Published online on: October 3, 2017 https://doi.org/10.3892/ol.2017.7132
Pages: 7091-7098
Copyright: © Jiang et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

Metrics: Total Views: 0 (Spandidos Publications: | PMC Statistics: )

Metrics: Total PDF Downloads: 0 (Spandidos Publications: | PMC Statistics: )

Cited By (CrossRef): 0 citations Loading Articles...

Abstract

Acute promyelocytic leukemia (APL) is characterized by the presence of the promyelocytic leukemia (PML)‑retinoic acid receptor‑α (RAR‑α) fusion protein. PML‑RARα can be cleaved by neutrophil elastase (NE) in several positions in cells in the promyelocytic stage, nuclear location signal (NLS)‑negative PML and NLS‑RARα may be the products of PML‑RARα by NE. The function of NLS‑RARα may be affected by the addition of NLS, which would alter its localization in cells, as the role of NLS is to identify proteins for transport to the nucleus. Preliminary experiments demonstrated that the overexpression of NLS‑RARα in HL‑60 cells could promote cellular proliferation and inhibit cellular differentiation. Following treatment with all‑trans retinoic acid (ATRA), the degree of cellular differentiation was enhanced. In the present study, the localization of NLS‑RARα was identified and its activity as a novel transcriptional factor was assessed, which may be critical in the development of APL. The location of NLS‑RARα was detected in the nucleus and cytoplasm by indirect immunofluorescence and western blot analysis, with expression in the nucleus revealed to be increased compared with that in the cytoplasm. Next, native‑PAGE was performed and NLS‑RARα and RXRα were revealed to form heterodimers in the nucleus. In addition, co‑immunoprecipitation revealed an interaction between NLS‑RARα and retinoid X receptor‑α (RXRα). An electrophoresis mobility shift assay (EMSA) indicated that NLS‑RARα could bind retinoic acid response elements (RAREs) in the presence of ATRA. Indeed, NLS‑RARα could bind RAREs just as WTRARα could, including the RAREs direct repeat‑2 (DR‑2) and DR‑5. In addition, results from a luciferase reporter gene assay demonstrated that NLS‑RARα could mediate the activity of RAREs that it bound. Together, these results indicated that NLS‑RARα may be a novel transcription factor that contributes to leukemogenesis by competitively binding RAREs as heterodimers with RXRα, just as PML‑RARα does, thus repressing the gene transcription essential for myeloid differentiation. These findings indicate the potential role of NLS‑RARα targeted therapy in APL.

View Figures

View References

1	de Thé H, Le Bras M and Lallemand-Breitenbach V: The cell biology of disease: Acute promyelocytic leukemia, arsenic, and PML bodies. J Cell Biol. 198:11–21. 2012. View Article : Google Scholar : PubMed/NCBI
2	Zelent A, Guidez F, Melnick A, Waxman S and Licht JD: Translocations of the RARalpha gene in acute promyelocytic leukemia. Oncogene. 20:7186–7203. 2001. View Article : Google Scholar : PubMed/NCBI
3	Melnick A and Licht JD: Deconstructing a disease: RARalpha, its fusion partners and their roles in the pathogenesis of acute promyelocytic leukemia. Blood. 93:3167–3215. 1999.PubMed/NCBI
4	Chang KS, Stass SA, Chu DT, Deaven LL, Trujillo JM and Freireich EJ: Characterization of a fusion cDNA (RARA/myl) transcribed from the t(15;17) translocation breakpoint in acute promyelocytic leukemia. Mol Cell Biol. 12:800–810. 1992. View Article : Google Scholar : PubMed/NCBI
5	Zhou GB, Chen SJ and Chen Z: Acute promyelocytic leukemia: A model of molecular target based therapy. Hematology. 10 Suppl 1:S270–S280. 2005. View Article : Google Scholar
6	Raelson JV, Nervi C, Rosenauer A, Benedetti L, Monczak Y, Pearson M, Pelicci PG and Miller WH Jr: The PML/RAR alpha Oncoprotein is a direct molecular target of retinoic acidin acute promyelocytic leukemia cells. Blood. 88:2826–2832. 1996.PubMed/NCBI
7	Zhou J, Pérès L, Honoré N, Nasr R, Zhu J and de Thé H: Dimerization-induced corepressor binding and relaxed DNA-binding specificity are critical for PML/RARA-induced immortalization. Proc Natl Acad Sci USA. 103:pp. 9238–9243. 2006, View Article : Google Scholar : PubMed/NCBI
8	Kamashev D, Vitoux D and De Thé H: PML-RARA-RXR oligomers mediate retinoid and rexinoid/cAMP cross-talk in acute promyelocytic leukemia cell differentiation. J Exp Med. 199:1163–1174. 2004. View Article : Google Scholar : PubMed/NCBI
9	Zhu J, Nasr R, Pérès L, Riaucoux-Lormière F, Honoré N, Berthier C, Kamashev D, Zhou J, Vitoux D, Lavau C and de Thé H: RXR is an essential component of the oncogenic PML/RARA complex in vivo. Cancer Cell. 12:23–35. 2007. View Article : Google Scholar : PubMed/NCBI
10	Vitaliano-Prunier A, Halftermeyer J, Ablain J, de Reynies A, Peres L, Le Bras M, Metzger D and de Thé H: Clearance of PML/RARA-bound promoters suffice to initiate APL differentiation. Blood. 124:3772–3780.. 2014. View Article : Google Scholar : PubMed/NCBI
11	Ablain J and de The H: Revisiting the differentiation paradigm in acute promyelocytic leukemia. Blood. 117:5795–5802. 2011. View Article : Google Scholar : PubMed/NCBI
12	Huang Y, Qiu J, Chen G and Dong S: Coiled-coil domain of PML is essential for the aberrant dynamics of PML-RARalpha, resulting in sequestration and decreased mobility of SMRT. Biochem Biophys Res Commun. 365:258–265. 2008. View Article : Google Scholar : PubMed/NCBI
13	de Thé H and Chen Z: Acute promyelocytic leukemia: Novel insights into the mechanisms of cure. Nat Rev Cancer. 10:775–783. 2010. View Article : Google Scholar : PubMed/NCBI
14	Ablain J and de Thé H: Retinoic acid signaling in cancer: The parable of acute promyelocytic leukemia. Int J Cancer. 135:2262–2272. 2014. View Article : Google Scholar : PubMed/NCBI
15	Lane AA and Ley TJ: Neutrophil elastase cleaves PML-RARalpha and is important for the development of acute promyelocytic leukemia in mice. 115:305–318. 2003.
16	Hayakawa F and Privalsky ML: Phosphorylation of PML by mitogen-activated protein kinases plays a key role in arsenic trioxide-mediated apoptosis. Cancer Cell. 5:389–401. 2004. View Article : Google Scholar : PubMed/NCBI
17	Cokol M, Nair R and Rost B: Finding nuclear localization signals. EMBO Rep. 1:411–415. 2000. View Article : Google Scholar : PubMed/NCBI
18	Zhang XW, Yan XJ, Zhou ZR, Yang FF, Wu ZY, Sun HB, Liang WX, Song AX, Lallemand-Breitenbach V, Jeanne M, et al: Arsenic trioxide controls the fate of the PML-RARalpha oncoprotein by directly binding PML. Science. 328:240–243.. 2010. View Article : Google Scholar : PubMed/NCBI
19	Wu TT, Chen C, Chen SM, Xu Y, Wang Y, Chen Z, Wang F, Xiao BK and Tao ZZ: Nuclear translocation of telomerase reverse transcriptase is a critical process in lymphatic metastasis of nasopharyngeal carcinoma. Oncol Lett. 9:265–269. 2015.PubMed/NCBI
20	Sánchez-Quesada C, López-Biedma A and Gaforio JJ: The differential localization of a methyl group confers a different anti-breast cancer activity to two triterpenes present in olives. Food Funct. 6:249–256. 2015. View Article : Google Scholar : PubMed/NCBI
21	Altucci L, Leibowitz MD, Ogilvie KM, de Lera AR and Gronemeyer H: RAR and RXR modulation in cancer and metabolic disease. Nat Rev Drug Discov. 6:793–810. 2007. View Article : Google Scholar : PubMed/NCBI
22	McCulloch D, Brown C and Iland H: Retinoic acid and arsenic trioxide in the treatment of acute promyelocytic leukemia: Current perspectives. Onco Targets Ther. 10:1585–1601. 2017. View Article : Google Scholar : PubMed/NCBI
23	Kakizuka A, Miller WH Jr, Umesono K, Warrell RP Jr, Frankel SR, Murty VV, Dmitrovsky E and Evans RM: Chromosomal translocation t(15;17) in human acute promyelocytic leukemia fuses RAR alpha with a novel putative transcription factor, PML. Cell. 66:663–674. 1991. View Article : Google Scholar : PubMed/NCBI
24	Choudhry A and DeLoughery TG: Bleeding and thrombosis in acute promyelocytic leukemia. Am J Hematol. 87:596–603. 2012. View Article : Google Scholar : PubMed/NCBI
25	Huang ME, Ye YC, Chen SR, Chai JR, Lu JX, Zhoa L, Gu LJ and Wang ZY: Use of all trans retinoic acid in the treatment of acute promyelocytic leukemia. Blood. 72:567–572. 1988.PubMed/NCBI
26	Hu XX, Zhong L, Zhang X, Gao YM and Liu BZ: NLS-RARα promotes proliferation and inhibits differentiation in HL-60 cells. Int J Med Sci. 11:247–254. 2014. View Article : Google Scholar : PubMed/NCBI
27	de Thé H, Lavau C, Marchio A, Chomienne C, Degos L and Dejean A: The PML-RAR alpha fusion mRNA generated by the t(15;17) translocation in acute promyelocytic leukemia encodes a functionally altered RAR. Cell. 66:675–684. 1991. View Article : Google Scholar : PubMed/NCBI
28	Zelent A, Guidez F, Melnick A, Waxman S and Licht JD: Translocations of the RARalpha gene in acute promyelocytic leukemia. Oncogene. 20:7186–7203. 2001. View Article : Google Scholar : PubMed/NCBI
29	Bastien J and Rochette-Egly C: Nuclear retinoid receptors and the transcription of retinoid-target genes. Gene. 328:1–16. 2004. View Article : Google Scholar : PubMed/NCBI
30	Planelles L, Medema JP, Hahne M and Hardenberg G: The expanding role of APRIL in cancer and immunity. Curr Mol Med. 8:829–844. 2008. View Article : Google Scholar : PubMed/NCBI
31	Delacroix L, Moutier E, Altobelli G, Legras S, Poch O, Choukrallah MA, Bertin I, Jost B and Davidson I: Cell-specific interaction of retinoic acid receptors with target genes in mouse embryonic fibroblasts and embryonic stem cells. Mol Cell Biol. 30:231–244. 2010. View Article : Google Scholar : PubMed/NCBI