Functions for fission yeast splicing factors SpSlu7 and SpPrp18 in alternative splice-site choice and stress-specific regulated splicing

PLoS One. 2017 Dec 13;12(12):e0188159. doi: 10.1371/journal.pone.0188159. eCollection 2017.

Abstract

Budding yeast spliceosomal factors ScSlu7 and ScPrp18 interact and mediate intron 3'ss choice during second step pre-mRNA splicing. The fission yeast genome with abundant multi-intronic transcripts, degenerate splice signals and SR proteins is an apt unicellular fungal model to deduce roles for core spliceosomal factors in alternative splice-site choice, intron retention and to study the cellular implications of regulated splicing. From our custom microarray data we deduce a stringent reproducible subset of S. pombe alternative events. We examined the role of factors SpSlu7 or SpPrp18 for these splice events and investigated the relationship to growth phase and stress. Wild-type log and stationary phase cells showed ats1+ exon 3 skipped and intron 3 retained transcripts. Interestingly the non-consensus 5'ss in ats1+ intron 3 caused SpSlu7 and SpPrp18 dependent intron retention. We validated the use of an alternative 5'ss in dtd1+ intron 1 and of an upstream alternative 3'ss in DUF3074 intron 1. The dtd1+ intron 1 non-canonical 5'ss yielded an alternative mRNA whose levels increased in stationary phase. Utilization of dtd1+ intron 1 sub-optimal 5' ss required functional SpPrp18 and SpSlu7 while compromise in SpSlu7 function alone hampered the selection of the DUF3074 intron 1 non canonical 3'ss. We analysed the relative abundance of these splice isoforms during mild thermal, oxidative and heavy metal stress and found stress-specific splice patterns for ats1+ and DUF3074 intron 1 some of which were SpSlu7 and SpPrp18 dependent. By studying ats1+ splice isoforms during compromised transcription elongation rates in wild-type, spslu7-2 and spprp18-5 mutant cells we found dynamic and intron context-specific effects in splice-site choice. Our work thus shows the combinatorial effects of splice site strength, core splicing factor functions and transcription elongation kinetics to dictate alternative splice patterns which in turn serve as an additional recourse of gene regulation in fission yeast.

Publication types

  • Validation Study

MeSH terms

  • Alternative Splicing*
  • Exons
  • Introns
  • Oligonucleotide Array Sequence Analysis
  • RNA Splice Sites*
  • RNA Splicing Factors / physiology*
  • Schizosaccharomyces pombe Proteins / physiology*
  • Stress, Physiological / genetics*

Substances

  • RNA Splice Sites
  • RNA Splicing Factors
  • Schizosaccharomyces pombe Proteins

Grants and funding

This work was supported by Programme for Advanced Research in Biosciences and Bioengineering (DBT/BF/PRIns/2011-12/28.9.2012) (UV); Infrastructure support University Grant Commission Centre for Advanced Studies (UV). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.