Upregulation of SPS100 gene expression by an antisense RNA via a switch of mRNA isoforms with different stabilities

Nucleic Acids Res. 2017 Nov 2;45(19):11144-11158. doi: 10.1093/nar/gkx737.

Abstract

Pervasive transcription of genomes generates multiple classes of non-coding RNAs. One of these classes are stable long non-coding RNAs which overlap coding genes in antisense direction (asRNAs). The function of such asRNAs is not fully understood but several cases of antisense-dependent gene expression regulation affecting the overlapping genes have been demonstrated. Using high-throughput yeast genetics and a limited set of four growth conditions we previously reported a regulatory function for ∼25% of asRNAs, most of which repress the expression of the sense gene. To further explore the roles of asRNAs we tested more conditions and identified 15 conditionally antisense-regulated genes, 6 of which exhibited antisense-dependent enhancement of gene expression. We focused on the sporulation-specific gene SPS100, which becomes upregulated upon entry into starvation or sporulation as a function of the antisense transcript SUT169. We demonstrate that the antisense effect is mediated by its 3' intergenic region (3'-IGR) and that this regulation can be transferred to other genes. Genetic analysis revealed that SUT169 functions by changing the relative expression of SPS100 mRNA isoforms from a short and unstable transcript to a long and stable species. These results suggest a novel mechanism of antisense-dependent gene regulation via mRNA isoform switching.

MeSH terms

  • Gene Expression Regulation, Fungal*
  • Immunoblotting
  • Microscopy, Fluorescence
  • RNA Isoforms / genetics*
  • RNA Stability / genetics
  • RNA, Antisense / genetics*
  • Reverse Transcriptase Polymerase Chain Reaction
  • Saccharomyces cerevisiae / genetics
  • Saccharomyces cerevisiae / metabolism
  • Saccharomyces cerevisiae / physiology
  • Saccharomyces cerevisiae Proteins / genetics*
  • Saccharomyces cerevisiae Proteins / metabolism
  • Spores, Fungal / genetics
  • Spores, Fungal / metabolism
  • Time-Lapse Imaging / methods
  • Up-Regulation*

Substances

  • RNA Isoforms
  • RNA, Antisense
  • SPS100 protein, S cerevisiae
  • Saccharomyces cerevisiae Proteins