Identifying RNA splicing factors using IFT genes in Chlamydomonas reinhardtii

Huawen Lin, Zhengyan Zhang, Carlo Iomini, Susan K. Dutcher

Research output: Contribution to journalArticle

Abstract

Intraflagellar transport moves proteins in and out of flagella/cilia and it is essential for the assembly of these organelles. Using whole-genome sequencing, we identified splice site mutations in two IFT genes, IFT81 (fla9) and IFT121 (ift121-2), which lead to flagellar assembly defects in the unicellular green alga Chlamydomonas reinhardtii. The splicing defects in these ift mutants are partially corrected by mutations in two conserved spliceosome proteins, DGR14 and FRA10. We identified a dgr14 deletion mutant, which suppresses the 3′ splice site mutation in IFT81, and a frameshift mutant of FRA10, which suppresses the 5′ splice sitemutation in IFT121. Surprisingly, we found dgr14-1 and fra10 mutations suppress both splice site mutations. We suggest these two proteins are involved in facilitating splice site recognition/interaction; in their absence some splice site mutations are tolerated. Nonsense mutations in SMG1, which is involved in nonsense-mediated decay, lead to accumulation of aberrant transcripts and partial restoration of flagellar assembly in the ift mutants. The high density of introns and the conservation of noncore splicing factors, together with the ease of scoring the ift mutant phenotype, make Chlamydomonas an attractive organism to identify new proteins involved in splicing through suppressor screening.

Original languageEnglish (US)
Article number170211
JournalOpen Biology
Volume8
Issue number3
DOIs
StatePublished - Jan 1 2018
Externally publishedYes

Fingerprint

Chlamydomonas reinhardtii
Genes
RNA
Mutation
Defects
RNA Splice Sites
Proteins
Algae
Introns
Restoration
Conservation
Carrier Proteins
Screening
Spliceosomes
Chlamydomonas
Chlorophyta
Flagella
Cilia
Nonsense Codon
Organelles

Keywords

  • Chlamydomonas
  • IFT
  • NMD
  • RNA splicing
  • Whole-genome sequencing

ASJC Scopus subject areas

  • Neuroscience(all)
  • Immunology
  • Biochemistry, Genetics and Molecular Biology(all)

Cite this

Identifying RNA splicing factors using IFT genes in Chlamydomonas reinhardtii. / Lin, Huawen; Zhang, Zhengyan; Iomini, Carlo; Dutcher, Susan K.

In: Open Biology, Vol. 8, No. 3, 170211, 01.01.2018.

Research output: Contribution to journalArticle

Lin, Huawen ; Zhang, Zhengyan ; Iomini, Carlo ; Dutcher, Susan K. / Identifying RNA splicing factors using IFT genes in Chlamydomonas reinhardtii. In: Open Biology. 2018 ; Vol. 8, No. 3.
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