TFIIS-Dependent Non-coding Transcription Regulates Developmental Genome Rearrangements

PLoS Genet. 2015 Jul 15;11(7):e1005383. doi: 10.1371/journal.pgen.1005383. eCollection 2015 Jul.

Abstract

Because of their nuclear dimorphism, ciliates provide a unique opportunity to study the role of non-coding RNAs (ncRNAs) in the communication between germline and somatic lineages. In these unicellular eukaryotes, a new somatic nucleus develops at each sexual cycle from a copy of the zygotic (germline) nucleus, while the old somatic nucleus degenerates. In the ciliate Paramecium tetraurelia, the genome is massively rearranged during this process through the reproducible elimination of repeated sequences and the precise excision of over 45,000 short, single-copy Internal Eliminated Sequences (IESs). Different types of ncRNAs resulting from genome-wide transcription were shown to be involved in the epigenetic regulation of genome rearrangements. To understand how ncRNAs are produced from the entire genome, we have focused on a homolog of the TFIIS elongation factor, which regulates RNA polymerase II transcriptional pausing. Six TFIIS-paralogs, representing four distinct families, can be found in P. tetraurelia genome. Using RNA interference, we showed that TFIIS4, which encodes a development-specific TFIIS protein, is essential for the formation of a functional somatic genome. Molecular analyses and high-throughput DNA sequencing upon TFIIS4 RNAi demonstrated that TFIIS4 is involved in all kinds of genome rearrangements, including excision of ~48% of IESs. Localization of a GFP-TFIIS4 fusion revealed that TFIIS4 appears specifically in the new somatic nucleus at an early developmental stage, before IES excision. RT-PCR experiments showed that TFIIS4 is necessary for the synthesis of IES-containing non-coding transcripts. We propose that these IES+ transcripts originate from the developing somatic nucleus and serve as pairing substrates for germline-specific short RNAs that target elimination of their homologous sequences. Our study, therefore, connects the onset of zygotic non coding transcription to the control of genome plasticity in Paramecium, and establishes for the first time a specific role of TFIIS in non-coding transcription in eukaryotes.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Cell Lineage
  • Genome*
  • Germ Cells
  • High-Throughput Nucleotide Sequencing
  • Paramecium tetraurelia / genetics
  • RNA Polymerase II / genetics
  • RNA, Long Noncoding / genetics*
  • Transcription, Genetic*
  • Transcriptional Elongation Factors / genetics*

Substances

  • RNA, Long Noncoding
  • Transcriptional Elongation Factors
  • transcription factor S-II
  • RNA Polymerase II

Associated data

  • GEO/GSE64682
  • SRA/SRP047508
  • SRA/SRX1022957

Grants and funding

This work was supported by grants from the Polish National Science Centre (#N N303 808840 to JKN, http://www.ncn.gov.pl/), the Polish Ministry of Science and Higher Education (#IP2010 028470 to JKN, http://www.nauka.gov.pl/), the French National Research Agency (collaborative projects ANR-10-BLAN-1603 “GENOMAC” and ANR-12-BSV6-0017 “INFERNO”, http://www.agence-nationale-recherche.fr/), the ARC Foundation for Cancer Research (#SFI20121205487 to MB, http://www.fondation-arc.org/) and the Centre National de la Recherche Scientifique (CNRS; an ATIP Plus grant to SD, http://www.cnrs.fr/). KMO, CDW and MB received exchange grants from the CNRS and the Polish Academy of Sciences (http://www.pan.pl/) in the framework of the CNRS GDRE “Paramecium Genome Dynamics and Evolution”. KMO was supported by PhD fellowship from the School of Molecular Biology—Institute of Biochemistry and Biophysics, PAS (http://www.ibb.waw.pl/) and exchange visit grant from the European Science Foundation (http://www.esf.org/, “Frontiers of Functional Genomics” program). The project was carried out in the framework of the CNRS-supported PICS project “Epigenetic targeting of programmed DNA elimination in the ciliate Paramecium tetraurelia” and the European Science Foundation COST action BM1102 “Ciliates as model systems to study genome evolution, mechanisms of non-Mendelian inheritance, and their roles in environmental adaptation” (http://www.cost.eu). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.