Yeast nucleoporin mutants are defective in pre-tRNA splicing

We have screened nucleoporin mutants for the inhibition of tRNA splicing, which has previously been proposed to be coupled to transport. Strains mutant for Nup49p or Nup116p, or genetically depleted of Nup145p, strongly accumulated unspliced pre-tRNAs. Splicing was inhibited for all 10 families of intron-containing pre-tRNA, but no effects on 5' or 3' end processing were detected. Strains mutant for Nup133p or Nsp1p accumulated lower levels of several unspliced pre-tRNAs. In contrast, no accumulation of any pre-tRNA was observed in strains mutant for Nup1p, Nup85p, or Nup100p. Other RNA processing reactions tested, pre-rRNA processing, pre-mRNA splicing, and small nucleolar and small nuclear RNA synthesis, were not clearly affected for any nucleoporin mutant. These data provide evidence for a coupling between pre-tRNA splicing and nuclear-cytoplasmic transport. Mutation of NUP49, NUP116, or NUP145 has previously been shown to lead to nuclear poly(A)+ RNA accumulation, indicating that these nucleoporins play roles in the transport of more than one class of RNA.

[1]  J. Sambrook,et al.  Molecular Cloning: A Laboratory Manual , 2001 .

[2]  E. Hurt,et al.  Nuclear transport , 1997, Science.

[3]  Schlaich Nl,et al.  Analysis of nucleocytoplasmic transport and nuclear envelope structure in yeast disrupted for the gene encoding the nuclear pore protein Nup1p. , 1995 .

[4]  F. Bischoff,et al.  Human RanGTPase-activating protein RanGAP1 is a homologue of yeast Rna1p involved in mRNA processing and transport. , 1995, Proceedings of the National Academy of Sciences of the United States of America.

[5]  R. Wepf,et al.  A novel nuclear pore protein Nup133p with distinct roles in poly(A)+ RNA transport and nuclear pore distribution. , 1994, The EMBO journal.

[6]  E. Hurt,et al.  Nup145p is required for nuclear export of mRNA and binds homopolymeric RNA in vitro via a novel conserved motif , 1994, Cell.

[7]  B. Séraphin,et al.  The POP1 gene encodes a protein component common to the RNase MRP and RNase P ribonucleoproteins. , 1994, Genes & development.

[8]  J. Broach,et al.  TPD1 of Saccharomyces cerevisiae encodes a protein phosphatase 2C-like activity implicated in tRNA splicing and cell separation , 1994, Molecular and cellular biology.

[9]  W. Boelens,et al.  Nuclear export of different classes of RNA is mediated by specific factors , 1994, The Journal of cell biology.

[10]  R. Lempicki,et al.  Saccharomyces cerevisiae U14 small nuclear RNA has little secondary structure and appears to be produced by post-transcriptional processing. , 1994, The Journal of biological chemistry.

[11]  G. Blobel,et al.  A temperature-sensitive NUP116 null mutant forms a nuclear envelope seal over the yeast nuclear pore complex thereby blocking nucleocytoplasmic traffic , 1993, The Journal of cell biology.

[12]  E. Hurt,et al.  Analysis of nucleo-cytoplasmic transport in a thermosensitive mutant of nuclear pore protein NSP1. , 1993, European journal of cell biology.

[13]  L. Guarente Synthetic enhancement in gene interaction: a genetic tool come of age. , 1993, Trends in genetics : TIG.

[14]  A. Hopper,et al.  The Saccharomyces cerevisiae LOS1 gene involved in pre-tRNA splicing encodes a nuclear protein that behaves as a component of the nuclear matrix. , 1993, The Journal of biological chemistry.

[15]  D. Goldfarb,et al.  Regulation of RNA processing and transport by a nuclear guanine nucleotide release protein and members of the Ras superfamily. , 1993, The EMBO journal.

[16]  A. Lee,et al.  GSP1 and GSP2, genetic suppressors of the prp20-1 mutant in Saccharomyces cerevisiae: GTP-binding proteins involved in the maintenance of nuclear organization , 1993, Molecular and cellular biology.

[17]  G. Fink,et al.  NUP2, a novel yeast nucleoporin, has functional overlap with other proteins of the nuclear pore complex. , 1993, Molecular biology of the cell.

[18]  P. Grandi,et al.  A new subclass of nucleoporins that functionally interact with nuclear pore protein NSP1. , 1992, The EMBO journal.

[19]  E. Hurt,et al.  NSP1 depletion in yeast affects nuclear pore formation and nuclear accumulation. , 1992, European journal of cell biology.

[20]  G. Blobel,et al.  A new family of yeast nuclear pore complex proteins , 1992, The Journal of cell biology.

[21]  C. Peebles,et al.  PTA1, an essential gene of Saccharomyces cerevisiae affecting pre-tRNA processing , 1992, Molecular and cellular biology.

[22]  L. Kühn,et al.  Export of mRNA from microinjected nuclei of Xenopus laevis oocytes , 1992, The Journal of cell biology.

[23]  C. Cole,et al.  Isolation and characterization of RAT1: an essential gene of Saccharomyces cerevisiae required for the efficient nucleocytoplasmic trafficking of mRNA. , 1992, Genes & development.

[24]  Y. Zhao,et al.  A conditional yeast mutant deficient in mRNA transport from nucleus to cytoplasm. , 1992, Proceedings of the National Academy of Sciences of the United States of America.

[25]  M. Carmo-Fonseca,et al.  Human nucleoporin p62 and the essential yeast nuclear pore protein NSP1 show sequence homology and a similar domain organization. , 1991, European journal of cell biology.

[26]  D. Tollervey,et al.  The small nucleolar RNP protein NOP1 (fibrillarin) is required for pre‐rRNA processing in yeast. , 1991, The EMBO journal.

[27]  C. Peebles,et al.  In vivo pre-tRNA processing in Saccharomyces cerevisiae , 1991, Molecular and cellular biology.

[28]  R. Rauhut,et al.  Yeast tRNA-splicing endonuclease is a heterotrimeric enzyme. , 1990, The Journal of biological chemistry.

[29]  G. Fink,et al.  The NUP1 gene encodes an essential component of the yeast nuclear pore complex , 1990, Cell.

[30]  J. Broach,et al.  A general screen for mutant of Saccharomyces cerevisiae deficient in tRNA biosynthesis. , 1989, Genetics.

[31]  G. Blobel,et al.  Yeast nuclear envelope proteins cross react with an antibody against mammalian pore complex proteins , 1989, The Journal of cell biology.

[32]  A. Hopper,et al.  Isolation of a yeast gene involved in species-specific pre-tRNA processing , 1988, Molecular and cellular biology.

[33]  N. Tanner,et al.  Binding interactions between yeast tRNA ligase and a precursor transfer ribonucleic acid containing two photoreactive uridine analogues. , 1988, Biochemistry.

[34]  A. McDowall,et al.  Nucleolar and nuclear envelope proteins of the yeast Saccharomyces cerevisiae. , 1988, European journal of cell biology.

[35]  D. Tollervey A yeast small nuclear RNA is required for normal processing of pre‐ribosomal RNA. , 1987, The EMBO journal.

[36]  M. W. Clark,et al.  The subnuclear localization of tRNA ligase in yeast , 1987, The Journal of cell biology.

[37]  E. Phizicky,et al.  Saccharomyces cerevisiae tRNA ligase. Purification of the protein and isolation of the structural gene. , 1986, The Journal of biological chemistry.

[38]  M. Zasloff,et al.  tRNA nuclear transport: Defining the critical regions of human tRNAi met by point mutagenesis , 1985, Cell.

[39]  L. Hartwell,et al.  Genetic analysis of the mitotic transmission of minichromosomes , 1985, Cell.

[40]  R. Ogden,et al.  Transfer RNA splicing in Saccharomyces cerevisiae: defining the substrates. , 1984, Nucleic acids research.

[41]  M. Zasloff tRNA transport from the nucleus in a eukaryotic cell: carrier-mediated translocation process. , 1983, Proceedings of the National Academy of Sciences of the United States of America.

[42]  J. Abelson,et al.  Precise excision of intervening sequences from precursor tRNAs by a membrane-associated yeast endonuclease , 1983, Cell.

[43]  R. Cortese,et al.  Order and intracellular location of the events involved in the maturation of a spliced tRNA , 1980, Nature.

[44]  A. Hopper,et al.  Processing of intervening sequences: a new yeast mutant which fails to excise intervening sequences from precursor tRNAs , 1980, Cell.

[45]  J. Beckmann,et al.  Transcription and processing of intervening sequences in yeast tRNA genes , 1978, Cell.

[46]  A. Hopper,et al.  A yeast mutant which accumulates precursor tRNAs , 1978, Cell.

[47]  N. Severs,et al.  Nuclear pore formation and the cell cycle in Saccharomyces cerevisiae. , 1977, Experimental cell research.

[48]  K. Shiokawa,et al.  The role of cytoplasmic membranes in controlling the transport of nuclear messenger RNA and initiation of protein synthesis. , 1974, Proceedings of the National Academy of Sciences of the United States of America.

[49]  J. Warner,et al.  The cytoplasmic maturation of a ribosomal precursor ribonucleic acid in yeast. , 1973, The Journal of biological chemistry.

[50]  L. Hartwell,et al.  Temperature-sensitive Yeast Mutant Defective in Ribonucleic Acid Production , 1969, Journal of bacteriology.

[51]  E. Hurt,et al.  Analysis of nucleocytoplasmic transport and nuclear envelope structure in yeast disrupted for the gene encoding the nuclear pore protein Nup1p. , 1995, European journal of cell biology.

[52]  S. Westaway,et al.  Splicing of tRNA Precursors , 1995 .

[53]  E. Izaurralde,et al.  RNA Export , 1995, Cell.

[54]  Gerald R. Fink,et al.  Methods in Yeast Genetics: A Laboratory Course Manual , 1987 .

[55]  J. Broach,et al.  The Molecular biology of the yeast Saccharomyces : metabolism and gene expression , 1982 .

[56]  C. Guthrie,et al.  Organization and Expression of tRNA Genes in Saccharomyces cerevisiae , 1982 .