Mutation in the prp12+ gene encoding a homolog of SAP130/SF3b130 causes differential inhibition of pre-mRNA splicing and arrest of cell-cycle progression in Schizosaccharomyces pombe

RNA. 2001 May;7(5):671-81. doi: 10.1017/s1355838201001200.

Abstract

prp12-1 is one of the mutants defective in pre-mRNA splicing at a nonpermissive temperature in Schizosaccharomyces pombe. We found that the prp12+ gene encodes a protein highly homologous with a human splicing factor, SAP130/SF3b130, a subunit of a U2 snRNP-associated complex SF3b. Prp12p was shown to interact genetically with Prp10p that is a homolog of SAP155/SF3b155, another subunit in SF3b, suggesting that Prp12p is a functional homolog of human SAP130/SF3b130. Prp12p tagged with GFP is uniformly localized in the nuclear DNA region. In addition to pre-mRNA splicing defects, the prp12-1 mutant produced elongated cells, a typical phenotype of cell division cycle (cdc) mutants, suggesting a possible link between pre-mRNA splicing and cell-cycle progression. We examined kinetics of splicing defects in prp12-1 and several other prp mutants using northern blot hybridization and found that, among all the tested pre-mRNAs, only Tflld pre-mRNA with low splicing efficiency showed detectable splicing defects at the nonpermissive temperature in prp12-1. In addition, we found that other prp mutants with the cdc phenotype also showed differential splicing defects in tested pre-mRNAs at the nonpermissive temperature. On the other hand, prp mutants that do not exhibit the cdc phenotype showed a rapid and complete block of pre-mRNA splicing in all the tested pre-mRNAs at the nonpermissive temperature, indicating that prp mutants with weak splicing defects have a tendency to exhibit the cdc phenotype. These results suggest that the cdc phenotype in prp12-1 is caused by a selective reduction of spliced transcripts encoding a protein (or proteins) required for G2/M transition.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Animals
  • Cell Cycle
  • Cell Nucleus / metabolism
  • Cloning, Molecular
  • Fungal Proteins / genetics*
  • Fungal Proteins / metabolism
  • Fungal Proteins / physiology
  • Genes, Fungal / physiology*
  • Green Fluorescent Proteins
  • Humans
  • Introns
  • Kinetics
  • Luminescent Proteins / metabolism
  • Molecular Sequence Data
  • Mutagenesis
  • Phosphoproteins
  • RNA Precursors*
  • RNA Splicing Factors
  • RNA Splicing*
  • RNA, Fungal*
  • RNA-Binding Proteins*
  • Ribonucleoprotein, U2 Small Nuclear
  • Schizosaccharomyces / cytology
  • Schizosaccharomyces / genetics
  • Schizosaccharomyces pombe Proteins*
  • Temperature
  • Transcription Factor TFIID
  • Transcription Factors, TFII / metabolism

Substances

  • Fungal Proteins
  • Luminescent Proteins
  • Phosphoproteins
  • Prp12 protein, S pombe
  • RNA Precursors
  • RNA Splicing Factors
  • RNA, Fungal
  • RNA-Binding Proteins
  • Ribonucleoprotein, U2 Small Nuclear
  • SF3B1 protein, human
  • SF3B3 protein, human
  • Schizosaccharomyces pombe Proteins
  • Sf3b1 protein, mouse
  • Transcription Factor TFIID
  • Transcription Factors, TFII
  • Green Fluorescent Proteins