Single-cell RNA-sequencing reveals pre-meiotic X-chromosome dosage compensation in Drosophila testis

PLoS Genet. 2021 Aug 17;17(8):e1009728. doi: 10.1371/journal.pgen.1009728. eCollection 2021 Aug.

Abstract

Dosage compensation equalizes X-linked expression between XY males and XX females. In male fruit flies, expression levels of the X-chromosome are increased approximately two-fold to compensate for their single X chromosome. In testis, dosage compensation is thought to cease during meiosis; however, the timing and degree of the resulting transcriptional suppression is difficult to separate from global meiotic downregulation of each chromosome. To address this, we analyzed testis single-cell RNA-sequencing (scRNA-seq) data from two Drosophila melanogaster strains. We found evidence that the X chromosome is equally transcriptionally active as autosomes in somatic and pre-meiotic cells, and less transcriptionally active than autosomes in meiotic and post-meiotic cells. In cells experiencing dosage compensation, close proximity to MSL (male-specific lethal) chromatin entry sites (CES) correlates with increased X chromosome transcription. We found low or undetectable levels of germline expression of most msl genes, mle, roX1 and roX2 via scRNA-seq and RNA-FISH, and no evidence of germline nuclear roX1/2 localization. Our results suggest that, although dosage compensation occurs in somatic and pre-meiotic germ cells in Drosophila testis, there might be non-canonical factors involved in the dosage compensation mechanism. The single-cell expression patterns and enrichment statistics of detected genes can be explored interactively in our database: https://zhao.labapps.rockefeller.edu/gene-expr/.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Base Sequence / genetics
  • Checkpoint Kinase 2 / genetics
  • Chromatin / metabolism
  • Chromosomal Proteins, Non-Histone / genetics
  • DNA Helicases / genetics
  • Dosage Compensation, Genetic / genetics*
  • Drosophila Proteins / genetics
  • Genes, X-Linked
  • Germ Cells / metabolism
  • Male
  • Meiosis / genetics
  • Nuclear Proteins / genetics
  • RNA / genetics
  • Sequence Analysis, RNA / methods
  • Single-Cell Analysis / methods
  • Testis / metabolism*
  • Transcription Factors / genetics
  • Transcription, Genetic
  • X Chromosome / genetics*
  • X Chromosome / metabolism

Substances

  • Chromatin
  • Chromosomal Proteins, Non-Histone
  • Drosophila Proteins
  • Nuclear Proteins
  • Transcription Factors
  • RNA
  • Checkpoint Kinase 2
  • lok protein, Drosophila
  • DNA Helicases