Caspase-3‑resistant uncleavable form of acidic leucine-rich nuclear phosphoprotein 32B potentiates leukemic cell apoptosis

Li,Cai‑Xia; Shen,Shao‑Ming; Wang,Li‑Shun; Yu,Yun

doi:10.3892/mmr.2014.3035

Caspase-3‑resistant uncleavable form of acidic leucine-rich nuclear phosphoprotein 32B potentiates leukemic cell apoptosis

Authors:
- Cai‑Xia Li
- Shao‑Ming Shen
- Li‑Shun Wang
- Yun Yu
View Affiliations

Affiliations: Key Laboratory of Cell Differentiation and Apoptosis of Chinese Ministry of Education, Center of Molecular Medicine, Rui‑Jin Hospital, Shanghai Jiao‑Tong University School of Medicine, Shanghai 200025, P.R. China
Published online on: December 3, 2014 https://doi.org/10.3892/mmr.2014.3035
Pages: 2813-2818

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

One member of the highly conserved acidic leucine‑rich nuclear phosphoprotein 32 kDa (ANP32) family of proteins, ANP32B, is critical for normal development, as demonstrated by a study in ANP32B‑deficient mice. Another study indicated that ANP32B was a direct substrate of caspase‑3, and was primarily cleaved at the sequence Ala‑Glu‑Val‑Asp, following Asp‑163. To investigate the significance of ANP32B cleavage in apoptosis, leukemic U937T cell lines were generated with inducible expression of ANP32B(wild type; WT), the uncleavable mutant ANP32B(D163A) and the N‑terminal fragment ANP32B(1‑163). Notably, overexpression of ANP32B(WT) and ANP32B(D163A) moderately increased and significantly enhanced etoposide‑induced apoptosis and caspase‑3 activation, whereas expression of ANP32B(1‑163) produced no effect. Two hypotheses have been generated, which may explain the distinct roles of the various ANP32B forms: i) ANP32B(WT) and ANP32B(D163A) localize in the nucleus while ANP32B(1‑163) mainly resides in the cytosol; or ii) ANP32B(WT) and ANP32B(D163A), but not ANP32B(1‑163), inhibit the expression of the anti‑apoptotic protein Bcl‑2. Based on these observations, caspase‑3‑resistant uncleavable ANP32B(D163A) is hypothesized to be pro‑apoptotic in leukemic cells.

View Figures

View References

1	Fuchs Y and Steller H: Programmed cell death in animal development and disease. Cell. 147:742–758. 2011. View Article : Google Scholar : PubMed/NCBI
2	Thornberry NA and Lazebnik Y: Caspases: enemies within. Science. 281:1312–1316. 1998. View Article : Google Scholar : PubMed/NCBI
3	Utz PJ and Anderson P: Life and death decisions: regulation of apoptosis by proteolysis of signaling molecules. Cell Death Differ. 7:589–602. 2000. View Article : Google Scholar : PubMed/NCBI
4	Johnson CE and Kornbluth S: Caspase cleavage is not for everyone. Cell. 134:720–721. 2008. View Article : Google Scholar : PubMed/NCBI
5	Timmer JC and Salvesen GS: Caspase substrates. Cell Death Differ. 14:66–72. 2007. View Article : Google Scholar
6	Matilla A and Radrizzani M: The Anp32 family of proteins containing leucine-rich repeats. Cerebellum. 4:7–18. 2005. View Article : Google Scholar : PubMed/NCBI
7	Adegbola O and Pasternack GR: Phosphorylated retinoblastoma protein complexes with pp32 and inhibits pp32-mediated apoptosis. J Biol Chem. 280:15497–15502. 2005. View Article : Google Scholar : PubMed/NCBI
8	Hoffarth S, Zitzer A, Wiewrodt R, et al: pp32/PHAPI determines the apoptosis response of non-small-cell lung cancer. Cell Death Differ. 15:161–170. 2008. View Article : Google Scholar
9	Jiang X, Kim HE, Shu H, et al: Distinctive roles of PHAP proteins and prothymosin-alpha in a death regulatory pathway. Science. 299:223–226. 2003. View Article : Google Scholar : PubMed/NCBI
10	Pan W, da Graca LS, Shao Y, et al: PHAPI/pp32 suppresses tumorigenesis by stimulating apoptosis. J Biol Chem. 284:6946–6954. 2009. View Article : Google Scholar : PubMed/NCBI
11	Schafer ZT, Parrish AB, Wright KM, et al: Enhanced sensitivity to cytochrome c-induced apoptosis mediated by PHAPI in breast cancer cells. Cancer Res. 66:2210–2218. 2006. View Article : Google Scholar : PubMed/NCBI
12	Shen SM, Yu Y, Wu YL, et al: Downregulation of ANP32B, a novel substrate of caspase-3, enhances caspase-3 activation and apoptosis induction in myeloid leukemic cells. Carcinogenesis. 31:419–426. 2010. View Article : Google Scholar
13	Brody JR, Kadkol SS, Hauer MC, et al: pp32 reduction induces differentiation of TSU-Pr1 cells. Am J Pathol. 164:273–283. 2004. View Article : Google Scholar
14	Kular RK, Cvetanovic M, Silferd S, et al: Neuronal differentiation is regulated by leucine-rich acidic nuclear protein (LANP), a member of the inhibitor of histone acetyltransferase complex. J Biol Chem. 284:7783–7792. 2009. View Article : Google Scholar : PubMed/NCBI
15	Puente LG, Carrière JF, Kelly JF and Megeney LA: Comparative analysis of phosphoprotein-enriched myocyte proteomes reveals widespread alterations during differentiation. FEBS Lett. 574:138–144. 2004. View Article : Google Scholar : PubMed/NCBI
16	Yu Y, Shen SM, Zhang FF, et al: Acidic leucine-rich nuclear phosphoprotein 32 family member B (ANP32B) contributes to retinoic acid-induced differentiation of leukemic cells. Biochem Biophys Res Commun. 423:721–725. 2012. View Article : Google Scholar : PubMed/NCBI
17	Amasaki H, Ogawa M, Nagasao J, et al: Distributional changes of BrdU, PCNA, E2F1 and PAL31 molecules in developing murine palatal rugae. Ann Anat. 185:517–523. 2003. View Article : Google Scholar
18	Sun W, Hattori N, Mutai H, et al: PAL31, a nuclear protein required for progression to the S phase. Biochem Biophys Res Commun. 280:1048–1054. 2001. View Article : Google Scholar : PubMed/NCBI
19	Fukukawa C, Tanuma N, Okada T, et al: pp32/ I-1(PP2A) negatively regulates the Raf-1/MEK/ERK pathway. Cancer Lett. 226:155–160. 2005. View Article : Google Scholar : PubMed/NCBI
20	Opal P, Garcia JJ, McCall AE, et al: Generation and characterization of LANP/pp32 null mice. Mol Cell Biol. 24:3140–3149. 2004. View Article : Google Scholar : PubMed/NCBI
21	Reilly PT, Afzal S, Gorrini C, et al: Acidic nuclear phosphoprotein 32kDa (ANP32)B-deficient mouse reveals a hierarchy of ANP32 importance in mammalian development. Proc Natl Acad Sci USA. 108:10243–10248. 2011. View Article : Google Scholar : PubMed/NCBI
22	Boer J, Bonten-Surtel J and Grosveld G: Overexpression of the nucleoporin CAN/NUP214 induces growth arrest, nucleocytoplasmic transport defects, and apoptosis. Mol Cell Biol. 18:1236–1247. 1998.PubMed/NCBI
23	Boer J, Bonten-Surtel J and Grosveld G: Overexpression of the nucleoporin CAN/NUP214 induces growth arrest, nucleocytoplasmic transport defects, and apoptosis. Mol Cell Biol. 18:1236–1247. 1998.PubMed/NCBI
24	Matassa AA, Carpenter L, Biden TJ, et al: PKCdelta is required for mitochondrial-dependent apoptosis in salivary epithelial cells. J Biol Chem. 276:29719–29728. 2001. View Article : Google Scholar : PubMed/NCBI
25	Qi X and Mochly-Rosen D: The PKCdelta -Abl complex communicates ER stress to the mitochondria - an essential step in subsequent apoptosis. J Cell Sci. 121:804–813. 2008. View Article : Google Scholar : PubMed/NCBI
26	Song MG, Gao SM, Du KM, et al: Nanomolar concentration of NSC606985, a camptothecin analog, induces leukemic-cell apoptosis through protein kinase Cdelta-dependent mechanisms. Blood. 105:3714–3721. 2005. View Article : Google Scholar : PubMed/NCBI
27	Munemasa Y, Suzuki T, Aizawa K, et al: Promoter region-specific histone incorporation by the novel histone chaperone ANP32B and DNA-binding factor KLF5. Mol Cell Biol. 28:1171–1181. 2008. View Article : Google Scholar :
28	Yu Y, Wang LS, Shen SM, et al: Subcellular proteome analysis of camptothecin analogue NSC606985-treated acute myeloid leukemic cells. J Proteome Res. 6:3808–3818. 2007. View Article : Google Scholar : PubMed/NCBI
29	Hill MM, Adrain C, Duriez PJ, et al: Analysis of the composition, assembly kinetics and activity of native Apaf-1 apoptosomes. EMBO J. 23:2134–2145. 2004. View Article : Google Scholar : PubMed/NCBI
30	Mutai H, Toyoshima Y, Sun W, et al: PAL31, a novel nuclear protein, expressed in the developing brain. Biochem Biophys Res Commun. 274:427–433. 2000. View Article : Google Scholar : PubMed/NCBI
31	Karin M and Lin A: NF-kappaB at the crossroads of life and death. Nat Immunol. 3:221–227. 2002. View Article : Google Scholar : PubMed/NCBI