RNA helicase DEAD-box-5 is involved in R-loop dynamics of preimplantation embryos

作者：Hyeonji Lee, Dong Wook Han, Seonho Yoo, Ohbeom Kwon, Hyeonwoo La, Chanhyeok Park, Heeji Lee, Kiye Kang, Sang Jun Uhm, Hyuk Song, Jeong Tae, Youngsok Choi, Kwonho Hong · 发表于：Animal Bioscience · 年份：2024 · DOI：10.5713/ab.23.0401 · 被引用次数：6 · 研究领域：DNA Repair Mechanisms、Reproductive Biology and Fertility、Genomics and Chromatin Dynamics

OBJECTIVE: R-loops are DNA:RNA triplex hybrids, and their metabolism is tightly regulated by transcriptional regulation, DNA damage response, and chromatin structure dynamics. R-loop homeostasis is dynamically regulated and closely associated with gene transcription in mouse zygotes. However, the factors responsible for regulating these dynamic changes in the R-loops of fertilized mouse eggs have not yet been investigated. This study examined the functions of candidate factors that interact with R-loops during zygotic gene activation. METHODS: In this study, we used publicly available next-generation sequencing datasets, including low-input ribosome profiling analysis and polymerase II chromatin immunoprecipitation-sequencing (ChIP-seq), to identify potential regulators of R-loop dynamics in zygotes. These datasets were downloaded, reanalyzed, and compared with mass spectrometry data to identify candidate factors involved in regulating R-loop dynamics. To validate the functions of these candidate factors, we treated mouse zygotes with chemical inhibitors using in vitro fertilization. Immunofluorescence with an anti-R-loop antibody was then performed to quantify changes in R-loop metabolism. RESULTS: We identified DEAD-box-5 (DDX5) and histone deacetylase-2 (HDAC2) as candidates that potentially regulate R-loop metabolism in oocytes, zygotes and two-cell embryos based on change of their gene translation. Our analysis revealed that the DDX5 inhibition of activity led to decreas...