Lactate Dehydrogenase A influences R-loop landscape

Using Cleavage Under Targets and Tagmentation-sequencing, we demonstrated that LDHA knockout disrupted R-loop homeostasis, altering S9.6-enriched peaks. Notably, exogenous lactate failed to rescue R-loop perturbations, suggesting LDHA's function extends beyond canonical metabolic activity. Furthermore, analysis of genomic binding sites of LDHA revealed a partial overlap between LDHA binding and R-loop-prone regions, suggesting that LDHA may regulate R-loop formation through direct binding. Transcriptomic analysis of 4T1 cells revealed widespread gene expression changes upon LDHA loss, yet only four genes correlated with both LDHA binding and R-loop peaks, indicating R-loops are secondary to LDHA's transcriptional regulation.