|
RNA helicase DEAD-box-5 is involved in R-loop dynamics of preimplantation embryos |
Hyeonji Lee1,a 
, Dong Wook Han2,a
, Seonho Yoo1
, Ohbeom Kwon1
, Hyeonwoo La1
, Chanhyeok Park1
, Heeji Lee1
, Kiye Kang1
, Sang Jun Uhm3
, Hyuk Song1
, Jeong Tae Do1
, Youngsok Choi1
, Kwonho Hong1,*
|
1Department of Stem Cell and Regenerative Biotechnology, Institute of Advanced Regenerative Science, Konkuk University, Seoul 05029, Korea
2Guangdong Provincial Key Laboratory of Large Animal Models for Biomedicine, Wuyi University, Jiangmen 529020, China
3Department of Animal Science, Sangji University, Wonju 26339, Korea |
Correspondence:
Kwonho Hong, Tel: +82-2-450-0560, Email: hongk@konkuk.ac.kr
|
Received: 6 October 2023 • Revised: 9 November 2023 • Accepted: 7 December 2023
aThese authors contributed equally to this work. |
|
|
| Abstract |
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 decreased R-loop accumulation in pronuclei, indicating its involvement in regulating R-loop dynamics. However, the inhibition of histone deacetylase-2 activity did not significantly affect R-loop levels in pronuclei.
Conclusion
These findings suggest that dynamic changes in R-loops during mouse zygote development are likely regulated by RNA helicases, particularly DDX5, in conjunction with transcriptional processes. Our study provides compelling evidence for the involvement of these factors in regulating R-loop dynamics during early embryonic development. |
| Keywords:
DEAD-box-5 (DDX5); Gene Transcription; R-loop; Zygote |
|
PDF Links
Full text via DOI
Download Citation
