alternative splicing and nonsense-mediated decay regulate telomerase reverse transcriptase (tert) expression during virus-induced lymphomagenesis in vivo可变剪接和nonsense-mediated衰减调节端粒酶逆转录酶在体内便占据(叔)表达式.pdfVIP

Amor et al. BMC Cancer 2010, 10:571 /1471-2407/10/571 RESEARCH ARTICLE Open Access Alternative splicing and nonsense-mediated decay regulate telomerase reverse transcriptase (TERT) expression during virus-induced lymphomagenesis in vivo 1 1,2 1,2 2,3 1* 1,2 Souheila Amor , Sylvie Remy , Ginette Dambrine , Yves Le Vern , Denis Rasschaert , Sylvie Laurent Abstract Background: Telomerase activation, a critical step in cell immortalization and oncogenesis, is partly regulated by alternative splicing. In this study, we aimed to use the Marek’s disease virus (MDV) T-cell lymphoma model to evaluate TERT regulation by splicing during lymphomagenesis in vivo, from the start point to tumor establishment. Results: We first screened cDNA libraries from the chicken MDV lymphoma-derived MSB-1 T- cell line, which we compared with B (DT40) and hepatocyte (LMH) cell lines. The chTERT splicing pattern was cell line-specific, despite similar high levels of telomerase activity. We identified 27 alternative transcripts of chicken TERT (chTERT). Five were in-frame alternative transcripts without in vitro telomerase activity in the presence of viral or chicken telomerase RNA (vTR or chTR), unlike the full-length transcript. Nineteen of the 22 transcripts with a premature termination codon (PTC) harbored a PTC more than 50 nucleotides upstream from the 3’ splice junction, and were therefore predicted targets for nonsense-mediated decay (NMD). The major PTC-containing alternatively spliced form identified in MSB1 (ie10) was targeted to the NMD pathway, as demonstrated by UPF1 silencing. We then studied three splicing events separately, and the balance between in-frame alternative splice variants (d5f and d10f)