Prostate cancer is the most prevalent cancer in males in developed countries. Tumor suppressor candidate 3 (TUSC3) has been identified as a putative tumor suppressor gene in prostate cancer, though its function has not been characterized. TUSC3 shares homologies with the yeast oligosaccharyltransferase (OST) complex subunit Ost3p, suggesting a role in protein glycosylation. We provide evidence that TUSC3 is part of the OST complex and affects N-linked glycosylation in mammalian cells. Loss of TUSC3 expression in DU145 and PC3 prostate cancer cell lines leads to increased proliferation, migration and invasion as well as accelerated xenograft growth in a PTEN negative background. TUSC3 downregulation also affects endoplasmic reticulum (ER) structure and stress response, which results in increased Akt signaling. Together, our findings provide first mechanistic insight in TUSC3 function in prostate carcinogenesis in general and N-glycosylation in particular.

Clinical Institute of Pathology Medical University of Vienna Austria

Department of Histology and Embryology Faculty of Medicine Masaryk University Brno Czech Republic

Department of Obstetrics and Gynecology Molecular Oncology Group Medical University of Vienna Austria

Division of Oncology Department of Internal Medicine 1 and Comprehensive Cancer Center Medical University of Vienna Austria

Institute for Cancer Research Department of Medicine 1 and Comprehensive Cancer Center Medical University of Vienna Austria

Zobrazit více v PubMed

Jemal A. et al. Global cancer statistics. CA Cancer J. Clin. 61, 69–90 (2008). PubMed

Berger M. F. et al. The genomic complexity of primary human prostate cancer. Nature 470, 214–220 (2011). PubMed PMC

Taylor B. S. et al. Integrative genomic profiling of human prostate cancer. Cancer Cell 18, 11–22 (2011). PubMed PMC

Bergerheim U. S., Kunimi K., Collins V. P. & Ekman P. Deletion mapping of chromosomes 8, 10, and 16 in human prostatic carcinoma. Gene. Chromosome. Canc. 3, 215–220 (1991). PubMed

Bova G. S. et al. Physical mapping of chromosome 8p22 markers and their homozygous deletion in a metastatic prostate cancer. Genomics 35, 46–54 (1996). PubMed

MacGrogan D., Levy A., Bova G. S., Isaacs W. B. & Bookstein R. Structure and methylation-associated silencing of a gene within a homozygously deleted region of human chromosome band 8p22. Genomics 35, 55–65 (1996). PubMed

Levy A., Dang U. C. & Bookstein R. High-density screen of human tumor cell lines for homozygous deletions of loci on chromosome arm 8p. Gene. Chromosome. Canc. 24, 42–47 (1999). PubMed

Arbieva Z. H. et al. High-resolution physical map and transcript identification of a prostate cancer deletion interval on 8p22. Genome Res. 10, 244–257 (2000). PubMed PMC

Cooke S. L. et al. High-resolution array CGH clarifies events occurring on 8p in carcinogenesis. BMC Cancer 8, 288 (2008). PubMed PMC

Pribill I. et al. High frequency of allelic imbalance at regions of chromosome arm 8p in ovarian carcinoma. Cancer Genet. Cytogenet. 129, 23–29 (2001). PubMed

Pils D. et al. Five genes from chromosomal band 8p22 are significantly down-regulated in ovarian carcinoma: N33 and EFA6R have a potential impact on overall survival. Cancer 104, 2417–2429 (2005). PubMed

Pils D. et al. Methylation status of TUSC3 is a prognostic factor in ovarian cancer. Cancer 119, 946–954 (2013). PubMed

Vanhara P. et al. Loss of the oligosaccharyl transferase subunit TUSC3 promotes proliferation and migration of ovarian cancer cells. Int. J. Oncol. 42, 1383–1389 (2013). PubMed

Kelleher D. J. & Gilmore R. An evolving view of the eukaryotic oligosaccharyltransferase. Glycobiology 16, 47R–62R (2006). PubMed

Kelleher D. J., Karaoglu D., Mandon E. C. & Gilmore R. Oligosaccharyltransferase isoforms that contain different catalytic STT3 subunits have distinct enzymatic properties. Mol. Cell 12, 101–111 (2003). PubMed

Mohorko E., Glockshuber R. & Aebi M. Oligosaccharyltransferase: the central enzyme of N-linked protein glycosylation. J. Inherit. Metab. Dis. 34, 869–878 (2011). PubMed

Garshasbi M. et al. A defect in the TUSC3 gene is associated with autosomal recessive mental retardation. Am. J. Hum. Genet. 82, 1158–1164 (2008). PubMed PMC

Khan M. A. et al. A novel deletion mutation in the TUSC3 gene in a consanguineous Pakistani family with autosomal recessive nonsyndromic intellectual disability. BMC Med. Genet. 12, 56 (2011). PubMed PMC

Molinari F. et al. Oligosaccharyltransferase-subunit mutations in nonsyndromic mental retardation. Am. J. Hum. Genet. 82, 1150–1157 (2008). PubMed PMC

Garshasbi M. et al. A novel nonsense mutation in TUSC3 is responsible for non-syndromic autosomal recessive mental retardation in a consanguineous Iranian family. Am. J. Med. Genet. A 155A, 1976–1980 (2011). PubMed

Ohtsubo K. & Marth J. D. Glycosylation in cellular mechanisms of health and disease. Cell 126, 855–867 (2006). PubMed

Helenius A. & Aebi M. Intracellular functions of N-linked glycans. Science 291, 2364–2369 (2001). PubMed

Freeze H. H. Genetic defects in the human glycome. Nat. Rev. Genet. 7, 537–551 (2006). PubMed

Hakomori S. Glycosylation defining cancer malignancy: new wine in an old bottle. Proc. Natl. Acad. Sci. U S A 99, 10231–10233 (2002). PubMed PMC

Fang M. et al. The ER UDPase ENTPD5 promotes protein N-glycosylation, the Warburg effect, and proliferation in the PTEN pathway. Cell 143, 711–724 (2011). PubMed

Hetz C., Martinon F., Rodriguez D. & Glimcher L. H. The unfolded protein response: integrating stress signals through the stress sensor IRE1alpha. Physiol. Rev. 91, 1219–1243 (2011). PubMed

Ozcan L. & Tabas I. Role of endoplasmic reticulum stress in metabolic disease and other disorders. Annu. Rev. Med. 63, 317–328 (2012). PubMed PMC

Harding H. P., Calfon M., Urano F., Novoa I. & Ron D. Transcriptional and translational control in the Mammalian unfolded protein response. Annu. Rev. Cell. Dev. Biol. 18, 575–599 (2002). PubMed

Schulz B. L. et al. Oxidoreductase activity of oligosaccharyltransferase subunits Ost3p and Ost6p defines site-specific glycosylation efficiency. Proc. Natl. Acad. Sci. U S A 106, 11061–11066 (2009). PubMed PMC

Contessa J. N. et al. Molecular imaging of N-linked glycosylation suggests glycan biosynthesis is a novel target for cancer therapy. Clin. Cancer Res. 16, 3205–3214 (2010). PubMed PMC

Shen Z., Huang S., Fang M. & Wang X. ENTPD5, an endoplasmic reticulum UDPase, alleviates ER stress induced by protein overloading in AKT-activated cancer cells. Cold Spring Harb. Symp. Quant. Biol. 76, 217–223 (2011). PubMed

Fu Y. et al. Pten null prostate tumorigenesis and AKT activation are blocked by targeted knockout of ER chaperone GRP78/BiP in prostate epithelium. Proc. Natl. Acad. Sci. U S A 105, 19444–19449 (2008). PubMed PMC

Dennis J. W., Granovsky M. & Warren C. E. Glycoprotein glycosylation and cancer progression. Biochim. Biophys. Acta 1473, 21–34 (1999). PubMed

Fukuda M. Possible roles of tumor-associated carbohydrate antigens. Cancer Res. 56, 2237–2244 (1996). PubMed

Lau K. S. et al. Complex N-glycan number and degree of branching cooperate to regulate cell proliferation and differentiation. Cell 129, 123–134 (2007). PubMed

Karaoglu D., Kelleher D. J. & Gilmore R. Functional characterization of Ost3p. Loss of the 34-kD subunit of the Saccharomyces cerevisiae oligosaccharyltransferase results in biased underglycosylation of acceptor substrates. J. Cell Biol. 130, 567–577 (1995). PubMed PMC

Zhou H. & Clapham D. E. Mammalian MagT1 and TUSC3 are required for cellular magnesium uptake and vertebrate embryonic development. Proc. Natl. Acad. Sci. U S A 106, 15750–15755 (2009). PubMed PMC

Ruiz-Canada C., Kelleher D. J. & Gilmore R. Cotranslational and posttranslational N-glycosylation of polypeptides by distinct mammalian OST isoforms. Cell 136, 272–283 (2009). PubMed PMC

Sato T. et al. STT3B-dependent posttranslational N-glycosylation as a surveillance system for secretory protein. Mol. Cell 47, 99–110 (2012). PubMed

Goel H. L., Li J., Kogan S. & Languino L. R. Integrins in prostate cancer progression. Endocr. Relat. Cancer 15, 657–664 (2008). PubMed PMC

Kariya Y. & Gu J. N-glycosylation of ss4 integrin controls the adhesion and motility of keratinocytes. PLoS One 6, e27084 (2011). PubMed PMC

Gabius H. J. M. V. D. W., Andre S. & Villalobo A. Down-regulation of the Epidermal Growth Factor Receptor by Altering N-Glycosylation: Emerging Role of beta1,4-Galactosyltransferases. Anticancer Res. 32, 1565–1572 (2012). PubMed

Kekeeva T. V. et al. [Abberant methylation of p16, HIC1, N33 and GSTP1 genes in tumor epitelium and tumor-associated stromal cells of prostate cancer]. Mol. Biol. (Mosk) 41, 79–85 (2007). PubMed

Hirota M., Kitagaki M., Itagaki H. & Aiba S. Quantitative measurement of spliced XBP1 mRNA as an indicator of endoplasmic reticulum stress. J. Toxicol. Sci. 31, 149–156 (2006). PubMed

van Schadewijk A., van't Wout E. F., Stolk J. & Hiemstra P. S. A quantitative method for detection of spliced X-box binding protein-1 (XBP1) mRNA as a measure of endoplasmic reticulum (ER) stress. Cell Stress Chaperones 17, 275–279 (2012). PubMed PMC

Samali A., Fitzgerald U., Deegan S. & Gupta S. Methods for monitoring endoplasmic reticulum stress and the unfolded protein response. Int. J. Cell Biol. 2010, 830307 (2010). PubMed PMC

Streit M., Lex A., Kalkusch M., Zatloukal K. & Schmalstieg D. Caleydo: connecting pathways and gene expression. Bioinformatics 25, 2760–2761 (2009). PubMed PMC

TUSC3: functional duality of a cancer gene