Mutant allele of rna14 in fission yeast affects pre-mRNA splicing

J Genet. 2016 Jun;95(2):389-97. doi: 10.1007/s12041-016-0652-z.

Abstract

Spliceosome and 3'-end processing complexes are necessary for the precursor mRNA (pre-mRNA) maturation. Spliceosome complex removes noncoding introns, while 3'-end processing involves in cleavage and addition of poly(A) tails to the nascent transcript. Rna14 protein in budding yeast has been implicated in cleavage and polyadenylation of mRNA in the nucleus but their role in the pre-mRNA splicing has not been studied. Here, we report the isolation of a mutant allele of rna14 in fission yeast, Schizosaccharomyces pombe that exhibits reduction in protein level of Chk1 at the nonpermissive temperature, primarily due to the defects in posttranscriptional processing. Reverse transcriptase-polymerase chain reaction analysis reveals defective splicing of the chk1(+) transcript at the nonpermissive temperature. Apart from chk1(+), the splicing of some other genes were also found to be defective at the nonpermissive temperature suggesting that Rna14 might be involved in pre-mRNA splicing. Subsequently, genetic interaction of Rna14 with prp1 and physical interactions with Prp28 suggest that the Rna14 might be part of a larger protein complex responsible for the pre-mRNA maturation.

MeSH terms

  • Alleles
  • Amino Acid Sequence
  • Checkpoint Kinase 1 / genetics*
  • Checkpoint Kinase 1 / metabolism
  • DEAD-box RNA Helicases / genetics
  • DEAD-box RNA Helicases / metabolism
  • Exons
  • Gene Expression Regulation, Fungal*
  • Introns
  • Mutation
  • RNA Precursors / genetics*
  • RNA Precursors / metabolism
  • RNA Splicing*
  • RNA, Fungal / genetics*
  • RNA, Fungal / metabolism
  • RNA, Messenger / genetics
  • RNA, Messenger / metabolism
  • Reverse Transcriptase Polymerase Chain Reaction
  • Schizosaccharomyces / genetics*
  • Schizosaccharomyces / metabolism
  • Schizosaccharomyces pombe Proteins / genetics*
  • Schizosaccharomyces pombe Proteins / metabolism
  • Sequence Alignment
  • Spliceosomes / genetics
  • Spliceosomes / metabolism
  • Temperature
  • mRNA Cleavage and Polyadenylation Factors / genetics*
  • mRNA Cleavage and Polyadenylation Factors / metabolism

Substances

  • PRP1 protein, S pombe
  • RNA Precursors
  • RNA, Fungal
  • RNA, Messenger
  • Rna14 protein, S pombe
  • Schizosaccharomyces pombe Proteins
  • mRNA Cleavage and Polyadenylation Factors
  • Checkpoint Kinase 1
  • Chk1 protein, S pombe
  • DEAD-box RNA Helicases