Comparative single- cell analysis of transcriptional bursting reveals the role of genome organization in de novo transcript origination

• 成果类型: Article
• 署名作者: Lee, Unjin; Li, Cong; Langer, Christopher B.; Svetec, Nicolas; Zhao, Li
• 署名单位: Rockefeller University
• 刊物名称: PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA
• ISSN/ISSBN: 0027-8733
• 发表日期: 2025-05-06
• 关键词: drosophila-melanogaster; gene-expression; dosage compensation; EVOLUTION; kinetics; sperm; LEVEL

摘要

Spermatogenesis is a key developmental process underlying the origination of newly evolved genes. However, rapid cell type-specific transcriptomic divergence of the Drosophila germline has posed a significant technical barrier for comparative single- cell RNA- sequencing studies. By quantifying a surprisingly strong correlation between species- and cell type-specific divergence in three closely related Drosophila species, we apply a statistical procedure to identify a core set of 198 genes that are highly predictive of cell type identity while remaining robust to species- specific differences that span over 25 to 30 My of evolution. We then utilize cell type classifications based on the 198- gene set to show how transcriptional divergence in cell type increases throughout spermatogenic developmental time. After validating these cross- species cell type classifications using RNA fluorescence in situ hybridization and imaging, we then investigate the influence of genome organization on the molecular evolution of spermatogenesis vis- a- vis transcriptional bursting. We first show altering transcriptional burst size contributes to premeiotic transcription and altering bursting frequency contributes to postmeiotic expression. We then report global differences in autosomal vs. X chromosomal transcription may arise in a developmental stage preceding full testis organogenesis by showing evolutionarily conserved decreases in X- linked transcription bursting kinetics in all examined somatic and germline cell types. Finally, we provide evidence supporting the cultivator model of de novo gene origination by demonstrating how the appearance of newly evolved testis- specific transcripts potentially provides short- range regulation of neighboring genes' transcriptional bursting properties during key stages of spermatogenesis.