Autoregulation of expression of the yeast Dbp2p ‘DEAD‐box’ protein is mediated by sequences in the conserved DBP2 intron.

The human p68, Saccharomyces cerevisiae DBP2 and Schizosaccharomyces pombe dbp2 genes are closely related members of the ‘DEAD‐box’ RNA helicase superfamily. All three genes contain an intron at a conserved site in RNA helicase motif V. The S.cerevisiae intron is unusual both for its position near the 3′‐end of the open reading frame and for its size, 1001 nucleotides. We show here that precise deletion of the intron has no effect on cell viability but leads to an increase in Dbp2p protein expression. Inefficient splicing due to the size of the intron can not account for this difference because the intron is efficiently spliced in Dbp2p‐deficient cells. Instead, there is a reciprocal relationship between the amount of Dbp2p in the cell and the efficiency with which DBP2 intron‐containing genes are expressed. Inactive Dbp2p mutants are efficiently expressed from DBP2 intron‐containing plasmids, and fragments of the DBP2 intron confer Dbp2p‐responsiveness on heterologous reporter introns. This suggest that there is an intron‐mediated negative feedback loop regulating DBP2 expression, and provides a possible explanation for the retention of such an unusual intron in S.cerevisiae.

[1]  F. He,et al.  Identification of a novel component of the nonsense-mediated mRNA decay pathway by use of an interacting protein screen. , 1995, Genes & development.

[2]  Isabelle lost,et al.  mRNAs can be stabilized by DEAD-box proteins , 1994, Nature.

[3]  J. Vilardell,et al.  Regulation of splicing at an intermediate step in the formation of the spliceosome. , 1994, Genes & development.

[4]  I. Bozzoni,et al.  Two different snoRNAs are encoded in introns of amphibian and human L1 ribosomal protein genes. , 1993, Nucleic acids research.

[5]  B. Sollner-Webb,et al.  Novel intron-encoded small nucleolar RNAs , 1993, Cell.

[6]  M. Rosbash,et al.  Short artificial hairpins sequester splicing signals and inhibit yeast pre-mRNA splicing , 1993, Molecular and cellular biology.

[7]  S M Burgess,et al.  Beat the clock: paradigms for NTPases in the maintenance of biological fidelity. , 1993, Trends in biochemical sciences.

[8]  S. Friend,et al.  Screening patients for heterozygous p53 mutations using a functional assay in yeast , 1993, Nature genetics.

[9]  S. Peltz,et al.  Stabilization and ribosome association of unspliced pre-mRNAs in a yeast upf1- mutant. , 1993, Proceedings of the National Academy of Sciences of the United States of America.

[10]  I. Bozzoni,et al.  A novel small nucleolar RNA (U16) is encoded inside a ribosomal protein intron and originates by processing of the pre‐mRNA. , 1993, The EMBO journal.

[11]  M. Fournier,et al.  The nucleolar snRNAs: catching up with the spliceosomal snRNAs. , 1993, Trends in biochemical sciences.

[12]  D. Libri,et al.  Pre‐mRNA secondary structure and the regulation of splicing , 1993, BioEssays : news and reviews in molecular, cellular and developmental biology.

[13]  R. Iggo,et al.  Mammalian p53 can function as a transcription factor in yeast. , 1992, Nucleic acids research.

[14]  P Linder,et al.  D‐E‐A‐D protein family of putative RNA helicases , 1992, Molecular microbiology.

[15]  S. Peltz,et al.  The product of the yeast UPF1 gene is required for rapid turnover of mRNAs containing a premature translational termination codon. , 1991, Genes & development.

[16]  J. Steitz,et al.  An intact Box C sequence in the U3 snRNA is required for binding of fibrillarin, the protein common to the major family of nucleolar snRNPs. , 1991, The EMBO journal.

[17]  M. Labouesse,et al.  A family of low and high copy replicative, integrative and single‐stranded S. cerevisiae/E. coli shuttle vectors , 1991, Yeast.

[18]  J. Rossi,et al.  Unexpected point mutations activate cryptic 3' splice sites by perturbing a natural secondary structure within a yeast intron. , 1991, Genes & development.

[19]  F. Eng,et al.  Structural basis for the regulation of splicing of a yeast messenger RNA , 1991, Cell.

[20]  C. Guthrie,et al.  A cold-sensitive mRNA splicing mutant is a member of the RNA helicase gene family. , 1991, Genes & development.

[21]  T. Kadesch,et al.  Post-transcriptional regulation of the human liver/bone/kidney alkaline phosphatase gene. , 1991, The Journal of biological chemistry.

[22]  D. Lane,et al.  p68 RNA helicase: identification of a nucleolar form and cloning of related genes containing a conserved intron in yeasts , 1991, Molecular and cellular biology.

[23]  J. Beggs,et al.  A suppressor of a yeast splicing mutation (prp8-l) encodes a putative ATP-dependent RNA helicase , 1991, Nature.

[24]  J. Abelson,et al.  Requirement of the RNA helicase-like protein PRP22 for release of messenger RNA from spliceosomes , 1991, Nature.

[25]  J. Steitz,et al.  RNA splicing. Alive with DEAD proteins. , 1991, Nature.

[26]  R. Rothstein Targeting, disruption, replacement, and allele rescue: integrative DNA transformation in yeast. , 1991, Methods in enzymology.

[27]  Janina Maier,et al.  Guide to yeast genetics and molecular biology. , 1991, Methods in enzymology.

[28]  J. H. Chen,et al.  The yeast PRP2 protein, a putative RNA-dependent ATPase, shares extensive sequence homology with two other pre-mRNA splicing factors. , 1990, Nucleic acids research.

[29]  D. Lane,et al.  Identification of a putative RNA helicase in E.coli. , 1990, Nucleic acids research.

[30]  J. Abelson,et al.  PRP5: a helicase-like protein required for mRNA splicing in yeast. , 1990, Proceedings of the National Academy of Sciences of the United States of America.

[31]  C. Guthrie,et al.  A putative ATP binding protein influences the fidelity of branchpoint recognition in yeast splicing , 1990, Cell.

[32]  R. W. Davis,et al.  Translation initiation and ribosomal biogenesis: involvement of a putative rRNA helicase and RPL46. , 1990, Science.

[33]  R. Gattoni,et al.  The 216-nucleotide intron of the E1A pre-mRNA contains a hairpin structure that permits utilization of unusually distant branch acceptors , 1989, Molecular and cellular biology.

[34]  V. Blinov,et al.  Two related superfamilies of putative helicases involved in replication, recombination, repair and expression of DNA and RNA genomes. , 1989, Nucleic acids research.

[35]  M. Scheffner,et al.  RNA helicase activity associated with the human p68 protein , 1989, Nature.

[36]  R. Iggo,et al.  Nuclear protein p68 is an RNA‐dependent ATPase. , 1989, The EMBO journal.

[37]  I. Graham,et al.  Effects of RNA secondary structure on alternative splicing of Pre-mRNA: Is folding limited to a region behind the transcribing RNA polymerase? , 1988, Cell.

[38]  I. Bozzoni,et al.  The accumulation of mature RNA for the Xenopus laevis ribosomal protein L1 is controlled at the level of splicing and turnover of the precursor RNA. , 1987, The EMBO journal.

[39]  C. Jacq,et al.  A mitochondrial RNA maturase gene transferred to the yeast nucleus can control mitochondrial mRNA splicing , 1986, Cell.

[40]  M. Dabeva,et al.  Autogenous regulation of splicing of the transcript of a yeast ribosomal protein gene. , 1986, Proceedings of the National Academy of Sciences of the United States of America.

[41]  G. Walter,et al.  Monoclonal antibodies specific for the carboxy terminus of simian virus 40 large T antigen , 1984, Journal of virology.

[42]  M. Yaffe,et al.  Two nuclear mutations that block mitochondrial protein import in yeast. , 1984, Proceedings of the National Academy of Sciences of the United States of America.

[43]  L. Guarente,et al.  Heme regulates transcription of the CYC1 gene of S. cerevisiae via an upstream activation site , 1983, Cell.

[44]  Jeffrey H. Miller Experiments in molecular genetics , 1972 .