The Function of ALYREF on UAP56-Mediated R-Loop Resolution
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R-loops arise during transcription, and serve important roles including class switch recombination, protection of genes from DNA methylation at GC rich promoters, and promotion of transcription termination, but they can also cause replication stress and double-strand breaks if they are allowed to accumulate and are not resolved by the cell’s innate processes of R-loop resolution, which greatly contributes to genomic instability. Cells do have methods of resolving persistent R-loops, but the mechanism of how these DNA replication and repair factors prevent R-loop accumulation remains unclear. Mutations in the TREX complex cause accumulation of co-transcriptional R-loops, and components of TREX, UAP56, and ALYREF, are of particular interest. UAP56 depletion in humans has been linked to a strong genomic instability phenotype, demonstrated in part by preliminary unpublished data showing that depletion of UAP56 results in an increased level of R-loops in HeLa cells. ALYREF associates with UAP56, and has been found overexpressed in cancerous tissue. These implications prompt the exploration of the function of ALYREF on UAP56 mediated R-loop resolution activity. Our data provided proof that UAP56 has R-loop dissociation activity, emphasized that ALYREF had a direct effect on the DNA/RNA helicase and R-loop dissociation activities of UAP56, and explored the basis of interaction between the two proteins through ALYREF variants.