We successfully identified several MRs including SOX3, TNFAIP3, TRAFD1, POU3F3, STAT2, and PML that govern the expression of a large collection of genes.
To this end, we generated retroviruses expressing either the PML open reading frame (LNC PML) or the PML cDNA fused to the 5′UTR (LNC 5′UTR PML).
A total of 372 PML cases were analyzed; 30 patients were asymptomatic and 342 were symptomatic at PML diagnosis.
LNC 5′UTR PML was obtained by cloning the murine Pml 5′UTR sequence directly in front of human PML IV in the polylinker of LNCX.
Since DAXX recruits proteins like H3.3 to PML-nuclear bodies (PML-NBs 24,56, PML-NBs 24,56and been shown to regulate PTEN37, we propose thaveDAXX, H3.3, PML and PTEN may form a chromatin complex that regulates gene transcription.
(a) Predicted protein protein interactions between PTEN, TP53, PML, DAXX and MCRS1 using the human protein interaction map.
For expression in MEFs or progenitors, MSCV retroviral constructs were used, HA or CFP tags were in frame with 5′ PML coding sequence.
Target genes were assessed using quantitative real-time PCR Taqman primer probes set for Irf8, Cd209a, Zbtb46, Bcl11a, Spi1, SpiB, Cebpe, Gfi1, Cebpa, Id2, Batf3, H2-Ke6, H2-Dma, H2-Q8, Tapbp, Tap2, Pml, Csf3r, Tbp, and Gapdh (Applied Biosystems).
AML1-ETO ChIP-seq data for Kasumi-1 cells were obtained from ref. 26 (GEO accession: GSE65427), PML and RARα ChIP-seq data for NB4 and UPR9 cells were obtained from ref. 27 (GEO accession: GSE18886).
Cluster 1 genes (ZBTB1, PML, ZNF44, SIX1, BCL6, ZNF450) were down-regulated by V and involved in gene transcription, whereas cluster 2 genes (IL8, IL1A, PTGS2, DTR, TNFAIP3, CXCL3) were up-regulated and linked to inflammatory response and cell proliferation.
Loss of regulation in the recruitment and coordination of key genes contained in these structures (Trp53, Pml, Brca1, Blm, etc).
