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Alternative Messenger RNA Splicing of Autophagic Gene Beclin 1 in Human B-cell Acute Lymphoblastic Leukemia Cells

  • Niu, Yu-Na ;
  • Liu, Qing-Qing ;
  • Zhang, Su-Ping ;
  • Yuan, Na ;
  • Cao, Yan ;
  • Cai, Jin-Yang ;
  • Lin, Wei-Wei ;
  • Xu, Fei ;
  • Wang, Zhi-Jian ;
  • Chen, Bo ;
  • Wang, Jian-Rong
  • Published : 2014.03.01

Abstract

Beclin 1 is a key factor for initiation and regulation of autophagy, which is a cellular catabolic process involved in tumorigenesis. To investigate the role of alternative splicing of Beclin1 in the regulation of autophagy in leukemia cells, Beclin1 mRNA from 6 different types of cell lines and peripheral blood mononuclear cells from 2 healthy volunteers was reversely transcribed, subcloned, and screened for alternative splicing. New transcript variants were analyzed by DNA sequencing. A transcript variant of Beclin 1 gene carrying a deletion of exon 11, which encoded a C-terminal truncation of Beclin 1 isoform, was found. The alternative isoform was assessed by bioinformatics, immunoblotting and subcellular localization. The results showed that this variable transcript is generated by alternative 3' splicing, and its translational product displayed a reduced activity in induction of autophagy by starvation, indicating that the spliced isoform might function as a dominant negative modulator of autophagy. Our findings suggest that the alternative splicing of Beclin 1 might play important roles in leukemogenesis regulated by autophagy.

Keywords

Beclin 1;alternative splicing;autophagy;acute leukemia

References

  1. Zeng L, Wang S, He S, et al (2013). Anesthetic management of low birth weight infants undergoing surgery for congenital heart disease without cardiopulmonary bypass. Nan Fang Yi Ke Da Xue Xue Bao, 33, 1806-10 (in Chinese).
  2. Yang Z, Goronzy JJ, Weyand CM (2014). The glycolytic enzyme PFKFB3/phosphofructokinase regulates autophagy. Autophagy, 10, 382-3. https://doi.org/10.4161/auto.27345
  3. Yu M, Gou WF, Zhao S, et al (2013). Beclin 1 expression is an independent prognostic factor for gastric carcinomas. Tumour BioI, 34, 1071-83. https://doi.org/10.1007/s13277-013-0648-8
  4. Yue Z, Jin S, Yang C, et al (2003). Beclin 1, an autophagy gene essential for early embryonic development, is a haploinsufficient tumor suppressor. Proc Natl Acad Sci USA, 100, 15077-82. https://doi.org/10.1073/pnas.2436255100
  5. Zhang J, Manley JL (2013). Misregulation of Pre-mRNA Alternative Splicing in Cancer. Cancer Discov,
  6. Zhang J, Ney PA (2009). Autophagy-dependent and -independent mechanisms of mitochondrial clearance during reticulocyte maturation. Autophagy, 5, 1064-5. https://doi.org/10.4161/auto.5.7.9749
  7. Zhang MX, Xu YJ, Zhu MC, Yan F (2013). Overexpressed ostepontin-c as a potential biomarker for esophageal squamous cell carcinoma. Asian Pac J Cancer Prev, 14, 7315-9. https://doi.org/10.7314/APJCP.2013.14.12.7315
  8. Zhang Z, Xin D, Wang P, et al (2009). Noisy splicing, more than expression regulation, explains why some exons are subject to nonsense-mediated mRNA decay. BMC Biol, 7, 23. https://doi.org/10.1186/1741-7007-7-23
  9. Sztainberg Y, Kuperman Y, Issler O, et al (2009). A novel corticotropin-releasing factor receptor splice variant exhibits dominant negative activity: a putative link to stress-induced heart disease. FASEB J, 23, 2186-96. https://doi.org/10.1096/fj.08-128066
  10. Salsman J, Pinder J, Tse B, et al (2013). The translation initiation factor 3 subunit eIF3K interacts with PML and associates with PML nuclear bodies. Exp Cell Res, 319, 2554-65. https://doi.org/10.1016/j.yexcr.2013.09.001
  11. Sandoval H, Thiagarajan P, Dasgupta SK, et al (2008). Essential role for Nix in autophagic maturation of erythroid cells. Nature, 454, 232-5. https://doi.org/10.1038/nature07006
  12. Smith CW, Valcarcel J (2000). Alternative pre-mRNA splicing: the logic of combinatorial control. Trends Biochem Sci, 25, 381-8. https://doi.org/10.1016/S0968-0004(00)01604-2
  13. Tang JY, Lee JC, Hou MF, et al (2013). Alternative splicing for diseases, cancers, drugs, and databases. Scientific World Journal, 2013, 703568.
  14. Torgersen ML, Simonsen A (2013). Autophagy: Friend or foe in the treatment of fusion protein-associated leukemias? Autophagy, 9, 2175-7. https://doi.org/10.4161/auto.26559
  15. Vazquez CL, Colombo MI (2010). Beclin 1 modulates the anti-apoptotic activity of Bcl-2: insights from a pathogen infection system. Autophagy, 6, 177-8. https://doi.org/10.4161/auto.6.1.10743
  16. Watson AS, Mortensen M, Simon AK (2011). Autophagy in the pathogenesis of myelodysplastic syndrome and acute myeloid leukemia. Cell Cycle, 10, 1719-25. https://doi.org/10.4161/cc.10.11.15673
  17. Wu XY, Chen J, Cao QH, et al (2013). Beclin 1 activation enhances chemosensitivity and predicts a favorable outcome for primary duodenal adenocarcinoma. Tumour BioI, 34, 713-22. https://doi.org/10.1007/s13277-012-0599-5
  18. Xing S, Zhang Y, Li J, et al (2012). Beclin 1 knockdown inhibits autophagic activation and prevents the secondary neurodegenerative damage in the ipsilateral thalamus following focal cerebral infarction. Autophagy, 8, 63-76. https://doi.org/10.4161/auto.8.1.18217
  19. Nicotra G, Mercalli F, Peracchio C, et al (2010). Autophagyactive beclin-1 correlates with favourable clinical outcome in non-Hodgkin lymphomas. Mod Pathol, 23, 937-50. https://doi.org/10.1038/modpathol.2010.80
  20. Mortensen M, and Simon AK (2010b). Nonredundant role of Atg7 in mitochondrial clearance during erythroid development. Autophagy, 6, 423-5. https://doi.org/10.4161/auto.6.3.11528
  21. Mortensen M, Watson AS, and Simon AK (2011). Lack of autophagy in the hematopoietic system leads to loss of hematopoietic stem cell function and dysregulated myeloid proliferation. Autophagy, 7, 1069-70. https://doi.org/10.4161/auto.7.9.15886
  22. Nascimento-Ferreira I, Nobrega C, Vasconcelos-Ferreira A, et al (2013). Beclin 1 mitigates motor and neuropathological deficits in genetic mouse models of Machado-Joseph disease. Brain, 136, 2173-88. https://doi.org/10.1093/brain/awt144
  23. Ouyang DY, Xu LH, He XH, et al (2013). Autophagy is differentially induced in prostate cancer LNCaP, DU145 and PC-3 cells via distinct splicing profiles of ATG5. Autophagy, 9, 20-32. https://doi.org/10.4161/auto.22397
  24. Pan Q, Shai O, Lee LJ, et al (2008). Deep surveying of alternative splicing complexity in the human transcriptome by highthroughput sequencing. Nat Genet, 40, 1413-5. https://doi.org/10.1038/ng.259
  25. Parkhitko AA, Favorova OO, Henske EP (2013). Autophagy: mechanisms, regulation, and its role in tumorigenesis. Biochemistry, 78, 355-67.
  26. Patel S, Dinesh-Kumar SP (2008). Arabidopsis ATG6 is required to limit the pathogen-associated cell death response. Autophagy, 4, 20-7. https://doi.org/10.4161/auto.5056
  27. Pua HH, Dzhagalov I, Chuck M, et al (2007). A critical role for the autophagy gene Atg5 in T cell survival and proliferation. J Exp Med, 204, 25-31. https://doi.org/10.1084/jem.20061303
  28. Sabirzhanov B, Keifer J (2011). Cloning and characterization of glutamate receptor subunit 4 (GLUA4) and its alternatively spliced isoforms in turtle brain. J Mol Neurosci, 44, 159-72. https://doi.org/10.1007/s12031-010-9405-2
  29. Liang XH, Kleeman LK, Jiang HH, et al (1998). Protection against fatal Sindbis virus encephalitis by beclin, a novel Bcl-2-interacting protein. J Virol, 72, 8586-96.
  30. Kervestin S, Jacobson A (2012). NMD: a multifaceted response to premature translational termination. Nat Rev Mol Cell Bioi, 13, 700-12. https://doi.org/10.1038/nrm3454
  31. Kornblihtt AR, Schor IE, Allo M, et al (2013). Alternative splicing: a pivotal step between eukaryotic transcription and translation. Nat Rev Mol Cell BioI, 14, 153-65. https://doi.org/10.1038/nrm3525
  32. Kundu M, Lindsten T, Yang CY, et al (2008). Ulk1 plays a critical role in the autophagic clearance of mitochondria and ribosomes during reticulocyte maturation. Blood, 112, 1493-502. https://doi.org/10.1182/blood-2008-02-137398
  33. Liu Y, Schiff M, Czymmek K, et al (2005). Autophagy regulates programmed cell death during the plant innate immune response. Cell, 121, 567-77. https://doi.org/10.1016/j.cell.2005.03.007
  34. Lu C, Li JY, Ge Z, et al (2013). Par-4/THAP1 complex and Notch3 competitively regulated pre-mRNA splicing of CCAR1 and affected inversely the survival of T-cell acute lymphoblastic leukemia cells. Oncogene, 32, 5602-13. https://doi.org/10.1038/onc.2013.349
  35. Lucin KM, O'Brien CE, Bieri G, et al (2013). Microglial beclin 1 regulates retromer trafficking and phagoc:ytosis and is impaired in Alzheimer's disease. Neuron, 79, 873-86. https://doi.org/10.1016/j.neuron.2013.06.046
  36. Margariti A, Li H, Chen T, et al (2013). XBP1 mRNA splicing triggers an autophagic response in endothelial cells through BECLIN-1 transcriptional activation. J BioI Chem, 288, 859-72. https://doi.org/10.1074/jbc.M112.412783
  37. Mizushima N, and Yoshimori T (2007). How to interpret LC3 immunoblotting. Autophagy, 3, 542-5. https://doi.org/10.4161/auto.4600
  38. Mortensen M, Ferguson DJ, Edelmann M, et al (2010a). Loss of autophagy in erythroid cells leads to defective removal of mitochondria and severe anemia in vivo. Proc Natl Acad Sci USA, 107, 832-7. https://doi.org/10.1073/pnas.0913170107
  39. Huang W, Choi W, Hu W, et al (2012). Crystal structure and biochemical analyses reveal Beclin 1 as a novel membrane binding protein. Cell Res, 22, 473-89. https://doi.org/10.1038/cr.2012.24
  40. He C, Klionsky DJ (2009). Regulation mechanisms and signaling pathways of autophagy. Annu Rev Genet, 43, 67-93. https://doi.org/10.1146/annurev-genet-102808-114910
  41. Huang JJ, Li HR, Huang Y, et al (2010). Beclin 1 expression: a predictor of prognosis in patients with extranodal natural killer T-cell lymphoma, nasal type. Autophagy, 6, 777-83. https://doi.org/10.4161/auto.6.6.12784
  42. Huang SC, Chang CL, Wang PS, et al (2009). Enterovirus 71-induced autophagy detected in vitro and in vivo promotes viral replication. J Med Virol, 81, 1241-52. https://doi.org/10.1002/jmv.21502
  43. Isakson P, Bjoras M, Boe SO, and Simonsen A (2010). Autophagy contributes to therapy-induced degradation of the PML/RARA oncoprotein. Blood, 116, 2324-31. https://doi.org/10.1182/blood-2010-01-261040
  44. Itakura E, Kishi C, Inoue K, Mizushima N (2008). Beclin 1 forms two distinct phosphatidylinositol 3-kinase complexes with mammalian Atg14 and UVRAG. Mol Bioi Cell, 19, 5360-72. https://doi.org/10.1091/mbc.E08-01-0080
  45. Jeffries EP, Denq WI, Bartko JC, Trakselis MA (2013). Identification, quantification, and evolutionary analysis of a novel isoform of MCM9. Gene, 519, 41-9. https://doi.org/10.1016/j.gene.2013.01.054
  46. Jiang L, Huang S, Li W, et al (2012). Expression of autophagy and ER stress-related proteins in primary salivary adenoid cystic carcinoma. Pathol Res Pract, 208, 635-41. https://doi.org/10.1016/j.prp.2012.07.008
  47. Kang R, Livesey KM, Zeh HJ, 3rd, et al (2011a). HMGB1 as an autophagy sensor in oxidative stress. Autophagy, 7, 904-6. https://doi.org/10.4161/auto.7.8.15704
  48. Kang R, Zeh HJ, Lotze MT, Tang D (2011b). The Beclin 1 network regulates autophagy and apoptosis. Cell Death Differ, 18, 571-80. https://doi.org/10.1038/cdd.2010.191
  49. Karpathiou G, Sivridis E, Koukourakis M, et al (2013). Autophagy and Bcl-2/BNIP3 death regulatory pathway in non-small cell lung carcinomas. APMIS, 121, 592-604. https://doi.org/10.1111/apm.12026
  50. Batista EL Jr., Kantarci AI, Hasturk H, Van Dyke TE (2013). Alternative splicing generates a diacylglycerol kinase alpha (DGKalpha) transcript that acts as a dominant negative modulator of superoxide production in localized aggressive periodontitis. J Periodontol, [Epub ahead of print].
  51. Chang YF, Imam JS, Wilkinson MF (2007). The nonsensemediated decay RNA surveillance pathway. Annu Rev Biochem, 76, 51-74. https://doi.org/10.1146/annurev.biochem.76.050106.093909
  52. Elzinga BM, Nyhan MJ, Crowley LC, et al (2013). Induction of autophagy by Imatinib sequesters Bcr-Abl in autophagosomes and down-regulates Bcr-Abl protein. Am J Hematol, 88, 455-62. https://doi.org/10.1002/ajh.23428
  53. Essick EE, Wilson RM, Pimentel DR, et al (2013). Adiponectin modulates oxidative stress-induced autophagy in cardiomyocytes. PLoS One, 8, e68697. https://doi.org/10.1371/journal.pone.0068697
  54. Fields J, Dumaop W, Rockenstein E, et al (2013). Age-dependent molecular alterations in the autophagy pathway in HIVE patients and in a gp120 tg mouse model: reversal with beclin-1 gene transfer. J Neurovirol, 19, 89-101. https://doi.org/10.1007/s13365-012-0145-7
  55. Han J, Hou W, Lu C, et al (2013). Interaction between Her2 and Beclin-1 proteins underlies a new mechanism of reciprocal regulation. J BioI Chem, 288, 20315-25. https://doi.org/10.1074/jbc.M113.461350

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