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oai:figshare.com:article/33716590 2026-09-13T22:02:27Z category_13 portal_316 item_type_3 month_year_09_2026

Data Sheet 1_Loss of lactate dehydrogenase A disrupts R-loop homeostasis and induces DNA damage in breast cancer cells.zip Shangzhen Li (20112128) Cheng Xu (352754) Shuling Chen (621838) Guoping Yin (9461705) Dawei Yang (796641) Genetics breast cancer Cut&Tag genomic stability lactate dehydrogenase A R-loop transcription Introduction<p>R-loops, three-stranded nucleic acid structures composed of an RNA-DNA hybrid and displaced single-stranded DNA, are tightly regulated by single-stranded DNA-binding proteins. However, the role of lactate dehydrogenase A (LDHA), an atypical DNA-binding protein and key glycolytic enzyme, in R-loop regulation remains unclear.</p>Methods<p>Cleavage Under Targets and Tagmentation sequencing (CUT&Tag-seq) was utilized to map R-loop and LDHA genomic landscapes in wild-type (WT) and LDHA knockout (KO) 4T1 cells. RNA-seq was employed for transcriptomic profiling. Immunofluorescence was used to evaluate the co-localization of LDHA and R-loops. Western blotting was performed to assess the protein levels of LDHA, RNase H1, and γ-H2AX.</p>Results<p>LDHA knockout disrupted R-loop homeostasis, resulting in altered S9.6-enriched peaks, with 199 peaks increased and 81 decreased in 4T1 cells. Exogenous lactate failed to rescue these R-loop perturbations, suggesting that LDHA regulates R-loops through a mechanism beyond its metabolic activity. Genomic analysis revealed partial overlap between LDHA-binding sites and R-loop-prone regions, indicating a potential direct role of LDHA in R-loop formation. Transcriptomic profiling showed widespread gene expression changes following LDHA loss, although only four genes were associated with both LDHA binding and R-loop peaks. Importantly, LDHA knockout induced accumulation of R-loop foci and DNA damage, both of which were reversed by RNase H1 expression.</p>Discussion<p>These findings identify LDHA as a novel regulator of R-loops and suggest that it may influence R-loop homeostasis through direct DNA binding rather than solely through its enzymatic role in glycolysis. Further studies are needed to elucidate the molecular mechanism underlying LDHA-mediated R-loop regulation.</p> 2026-09-13T22:02:27Z Dataset Dataset 10.3389/fgene.2026.1942597.s001 https://figshare.com/articles/dataset/Data_Sheet_1_Loss_of_lactate_dehydrogenase_A_disrupts_R-loop_homeostasis_and_induces_DNA_damage_in_breast_cancer_cells_zip/33716590 CC BY 4.0