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Intron 5 of RUNX1 Gene Harbor a Putative Promoter Region.

The t(8:21) is one of the most frequent chromosomal translocation found in leukemia. To date all the break points mapped for this translocation, are located in intron 5 of RUNX1 gene. Interestingly, this intron exhibit structural chromatin elements like Topoisomerase II and DNase I hypersensitive sites, which have been wildly associated to presence of transcriptional cis regulatory elements. Therefore, we hypothesize that regulatory elements are harbored in intron5 of the RUNX1 gene. In order to test this hypothesis, we performed bioinformatics analysis of ChIPseq data available from the ENCODE database to identify regions of intron 5 enriched in epigenetics marks characteristic of regulatory elements. Our results shown that in hematopoietic cells there is one region highly enrich in K4H3me, K4H3me3 and K27H3ac that colocalizes with DNase I hypersensitive sites. These marks have been associated with promoters modules. Indeed, when we cloned this region in the pGL3 basic reporter vector, we found that it activates expression of the reporter gene luciferase in an orientation dependent manner. Using ETS databases we have assemble a putative messenger for this promoter. Moreover, by RT‐PCR we have shown that this RNA is expressed both in Jurkat and HL60 cells. Taken together our results demonstrate the presence of a putative promoter region located in intron 5 of RUNX1 gene that is active in hematopoietic cells.