A Dual Upstream Open Reading Frame-based Autoregulatory Circuit Controlling Polyamine-responsive Translation*

A novel form of translational regulation is described for the key polyamine biosynthetic enzyme S-adenosylmethionine decarboxylase (AdoMetDC). Plant AdoMetDC mRNA 5′ leaders contain two highly conserved overlapping upstream open reading frames (uORFs): the 5′ tiny and 3′ small uORFs. We demonstrate that the small uORF-encoded peptide is responsible for constitutively repressing downstream translation of the AdoMetDC proenzyme ORF in the absence of increased polyamine levels. This first example of a sequence-dependent uORF to be described in plants is also functional in Saccharomyces cerevisiae. The tiny uORF is required for normal polyamine-responsive AdoMetDC mRNA translation, and we propose that this is achieved by control of ribosomal recognition of the occluded small uORF, either by ribosomal leaky scanning or by programmed -1 frameshifting. In vitro expression demonstrated that both the tiny and the small uORFs are translated. This tiny/small uORF configuration is highly conserved from moss to Arabidopsis thaliana, and a more diverged tiny/small uORF arrangement is found in the AdoMetDC mRNA 5′ leader of the single-celled green alga Chlamydomonas reinhardtii, indicating an ancient origin for the uORFs.

[1]  J. F. Atkins,et al.  Autoregulatory frameshifting in decoding mammalian ornithine decarboxylase antizyme , 1995, Cell.

[2]  P. Coffino Ubiquitin and proteasomes: Regulation of cellular polyamines by antizyme , 2001, Nature Reviews Molecular Cell Biology.

[3]  A. Michael,et al.  Overexpression of arginine decarboxylase in transgenic plants. , 1997, The Biochemical journal.

[4]  P. Torrigiani,et al.  Characterization of monocot and dicot plant S-adenosyl-l-methionine decarboxylase gene families including identification in the mRNA of a highly conserved pair of upstream overlapping open reading frames. , 2001, The Biochemical journal.

[5]  Ali Nahvi,et al.  Genetic control by a metabolite binding mRNA. , 2002, Chemistry & biology.

[6]  Mark Gerstein,et al.  Revisiting the codon adaptation index from a whole-genome perspective: analyzing the relationship between gene expression and codon occurrence in yeast using a variety of models. , 2003, Nucleic acids research.

[7]  Jonathan D. G. Jones,et al.  Effective vectors for transformation, expression of heterologous genes, and assaying transposon excision in transgenic plants , 1992, Transgenic Research.

[8]  J. G. Georgatsos,et al.  Concentration-dependent effects of natural polyamines on peptide chain initiation and elongation in a cell-free system of protein synthesis , 1990, Molecular and Cellular Biochemistry.

[9]  K. Kashiwagi,et al.  Polyamines: mysterious modulators of cellular functions. , 2000, Biochemical and biophysical research communications.

[10]  M. Sachs,et al.  A nascent polypeptide domain that can regulate translation elongation , 2004, Proceedings of the National Academy of Sciences of the United States of America.

[11]  M. Mayer,et al.  Abrogation of Upstream Open Reading Frame-mediated Translational Control of a Plant S-Adenosylmethionine Decarboxylase Results in Polyamine Disruption and Growth Perturbations* , 2002, The Journal of Biological Chemistry.

[12]  L. Ryabova,et al.  Ribosome shunting in the cauliflower mosaic virus 35S RNA leader is a special case of reinitiation of translation functioning in plant and animal systems. , 2000, Genes & development.

[13]  R. Sharma,et al.  Polyamines inhibit the protein kinase 380--catalyzed phosphorylation of eukaryotic initiation factor 2 alpha. , 1982, Science.

[14]  L. Shantz,et al.  Role of the 5'-untranslated region of mRNA in the synthesis of S-adenosylmethionine decarboxylase and its regulation by spermine. , 1994, The Biochemical journal.

[15]  S. Clough,et al.  Floral dip: a simplified method for Agrobacterium-mediated transformation of Arabidopsis thaliana. , 1998, The Plant journal : for cell and molecular biology.

[16]  A. Kingsman,et al.  A retrovirus-like strategy for expression of a fusion protein encoded by yeast transposon Ty1 , 1985, Nature.

[17]  M. Kozak,et al.  Pushing the limits of the scanning mechanism for initiation of translation , 2002, Gene.

[18]  T. Henkin,et al.  The L box regulon: Lysine sensing by leader RNAs of bacterial lysine biosynthesis genes , 2003, Proceedings of the National Academy of Sciences of the United States of America.

[19]  Koreaki Ito,et al.  The Ribosomal Exit Tunnel Functions as a Discriminating Gate , 2002, Cell.

[20]  G. Mize,et al.  In Vitro Translation of the Upstream Open Reading Frame in the Mammalian mRNA EncodingS-Adenosylmethionine Decarboxylase* , 2000, The Journal of Biological Chemistry.

[21]  N. Sonenberg,et al.  Translational control of gene expression , 2000 .

[22]  R. Elble A simple and efficient procedure for transformation of yeasts. , 1992, BioTechniques.

[23]  J. R. Johnston Molecular genetics of yeast :a practical approach , 1994 .

[24]  A. Hinnebusch,et al.  Physical evidence for distinct mechanisms of translational control by upstream open reading frames , 2001, The EMBO journal.

[25]  F. Messenguy,et al.  A segment of mRNA encoding the leader peptide of the CPA1 gene confers repression by arginine on a heterologous yeast gene transcript , 1994, Molecular and cellular biology.

[26]  D. Morris,et al.  Upstream Open Reading Frames as Regulators of mRNA Translation , 2000, Molecular and Cellular Biology.

[27]  Hiroshi Kobayashi,et al.  Estimation of polyamine binding to macromolecules and ATP in bovine lymphocytes and rat liver. , 1991, The Journal of biological chemistry.

[28]  D. Morris,et al.  Polyamine regulation of ribosome pausing at the upstream open reading frame of S-adenosylmethionine decarboxylase. , 2001, The Journal of biological chemistry.

[29]  A. Geballe,et al.  Translational Effects of Mutations and Polymorphisms in a Repressive Upstream Open Reading Frame of the Human Cytomegalovirus UL4 Gene , 1999, Journal of Virology.

[30]  D. Morris,et al.  The Upstream Open Reading Frame of the mRNA Encoding S-Adenosylmethionine Decarboxylase Is a Polyamine-responsive Translational Control Element* , 1996, The Journal of Biological Chemistry.

[31]  Igor P. Ivanov,et al.  Antizyme expression: a subversion of triplet decoding, which is remarkably conserved by evolution, is a sensor for an autoregulatory circuit , 2000, Nucleic acids research.

[32]  Seymour S. Cohen A Guide to the Polyamines , 1998 .

[33]  Ronald R. Breaker,et al.  Thiamine derivatives bind messenger RNAs directly to regulate bacterial gene expression , 2002, Nature.

[34]  M. M. Bradford A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. , 1976, Analytical biochemistry.

[35]  A. Cigan,et al.  Yeast translation initiation suppressor sui2 encodes the alpha subunit of eukaryotic initiation factor 2 and shares sequence identity with the human alpha subunit. , 1989, Proceedings of the National Academy of Sciences of the United States of America.

[36]  A. Michael,et al.  Expression proteomics identifies biochemical adaptations and defense responses in transgenic plants with perturbed polyamine metabolism , 2004, FEBS letters.

[37]  Haibiao Gong,et al.  The Pivotal Roles of the Plant S-Adenosylmethionine Decarboxylase 5′ Untranslated Leader Sequence in Regulation of Gene Expression at the Transcriptional and Posttranscriptional Levels1 , 2005, Plant Physiology.

[38]  A. Geballe,et al.  Coding sequence-dependent ribosomal arrest at termination of translation , 1996, Molecular and cellular biology.

[39]  S. Blair Hedges,et al.  The origin and evolution of model organisms , 2002, Nature Reviews Genetics.

[40]  J. Hill,et al.  Cell-specific translation of S-adenosylmethionine decarboxylase mRNA. Regulation by the 5' transcript leader. , 1992, The Journal of biological chemistry.

[41]  Z. Wang,et al.  Evolutionarily conserved features of the arginine attenuator peptide provide the necessary requirements for its function in translational regulation. , 2000, The Journal of biological chemistry.