Drosophila Trf4-1 involves in mRNA and primary miRNA transcription
Yongxiang Liu1, Ming Wang2, Xin Liu3
1State Key Laboratory of Virology, College of Life Sciences, Wuhan University, Wuhan, Hubei, 430072, China; Laboratory of RNA Virology, Wuhan Institute of Virology, Chinese Academy of Sciences, Wuhan, Hubei, 430071, China.
Abstract:
Drosophila Trf4-1 (DmTrf4-1) is a polyadenylation polymerase or terminal nucleotidyl transferase (PAP/TENT) that has been reported to add poly(A) tails to snRNAs in nucleus or mRNAs in cytoplasm. Here, we found that the loss of Trf4-1 resulted in the reduction of mRNAs and primary miRNAs (pri-miRNAs) in both Drosophila S2 cells and adult flies. Interestingly, the role of Trf4-1 in transcription is independent of its PAP/TENT activity. Moreover, using the chromatin immunoprecipitation assay, we uncovered that the loss of Trf4-1 led to abnormal RNA polymerase II accumulation and reduced H3K4me3 binding in promoter regions. Thus, our study indicates a positive role of Trf4-1 in the transcription of mRNAs and pri-miRNAs.
Insights
Drosophila Trf4-1 protein plays a crucial role in gene transcription, independent of its polyadenylation activity. Its absence reduces mRNA and pri-miRNA levels by affecting RNA polymerase II and H3K4me3 binding.
Area of Science:
- Molecular Biology
- Genetics
- Biochemistry
Background:
- Drosophila Trf4-1 is a polyadenylation polymerase/terminal nucleotidyl transferase (PAP/TENT).
- It is known to add poly(A) tails to snRNAs and mRNAs.
- Its precise role in gene expression regulation requires further investigation.
Purpose of the Study:
- To investigate the function of Drosophila Trf4-1 (DmTrf4-1) in gene expression.
- To determine if DmTrf4-1's role in transcription is linked to its enzymatic activity.
- To elucidate the molecular mechanisms underlying DmTrf4-1's transcriptional role.
Main Methods:
- Analysis of mRNA and primary miRNA levels in Drosophila S2 cells and adult flies.
- Chromatin immunoprecipitation (ChIP) assays to assess protein binding and histone modifications.
- Investigating the impact of Trf4-1 loss on RNA polymerase II and H3K4me3 at promoter regions.
Main Results:
- Loss of Trf4-1 significantly reduced mRNA and primary miRNA levels.
- DmTrf4-1's role in transcription is independent of its PAP/TENT activity.
- Trf4-1 deficiency caused abnormal RNA polymerase II accumulation and decreased H3K4me3 binding at promoters.
Conclusions:
- Drosophila Trf4-1 positively regulates the transcription of mRNAs and pri-miRNAs.
- DmTrf4-1 influences transcription through mechanisms involving RNA polymerase II and H3K4me3 modification.
- The study reveals a novel, non-enzymatic role for Trf4-1 in gene transcription.
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