Posttranscriptional Regulation by the Upstream Open Reading Frame of the Phosphoethanolamine N-Methyltransferase Gene

Phosphoethanolamine N-methyltransferase (PEAMT) is involved in choline biosynthesis in plants. The 5′ untranslated region (UTR) of several PEAMT genes was found to contain an upstream open reading frame (uORF). We generated transgenic Arabidopsis calli that expressed a chimeric gene constructed by fusing the 5′ UTR of the Arabidopsis PEAMT gene (AtNMT1) upstream of the β-glucuronidase gene. The AtNMT1 uORF was found to be involved in declining levels of the chimeric gene mRNA and repression of downstream β-glucuronidase gene translation in the calli when the cells were treated with choline. Further, we discuss the role of the uORF.

[1]  Xiaoqun Wang,et al.  Silencing of Phosphoethanolamine N-Methyltransferase Results in Temperature-Sensitive Male Sterility and Salt Hypersensitivity in Arabidopsis Article, publication date, and citation information can be found at www.plantcell.org/cgi/doi/10.1105/tpc.001701. , 2002, The Plant Cell Online.

[2]  F. Dietrich,et al.  Identification and characterization of upstream open reading frames (uORF) in the 5′ untranslated regions (UTR) of genes in Saccharomyces cerevisiae , 2005, Current Genetics.

[3]  T. Nanmori,et al.  Similar regulation patterns of choline monooxygenase, phosphoethanolamine N-methyltransferase and S-adenosyl-L-methionine synthetase in leaves of the halophyte Atriplex nummularia L. , 2005, Plant & cell physiology.

[4]  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.

[5]  Sjef Smeekens,et al.  A Conserved Upstream Open Reading Frame Mediates Sucrose-Induced Repression of Translation , 2004, The Plant Cell Online.

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

[7]  C Saccone,et al.  Analysis of oligonucleotide AUG start codon context in eukariotic mRNAs. , 2000, Gene.

[8]  E. Weretilnyk,et al.  Phosphocholine synthesis in spinach : Characterization of phosphoethanolamine N-methyltransferase , 2000 .

[9]  S. Coughlan,et al.  Glycinebetaine biosynthesis and its control in detached secondary leaves of spinach , 1982, Planta.

[10]  C. Rodrigues-Pousada,et al.  Post‐termination ribosome interactions with the 5′UTR modulate yeast mRNA stability , 1999, The EMBO journal.

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

[12]  C. Curie,et al.  A protocol for transient gene expression in Arabidopsis thaliana protoplasts isolated from cell suspension cultures , 1992 .

[13]  A. Hanson,et al.  Enhanced synthesis of choline and glycine betaine in transgenic tobacco plants that overexpress phosphoethanolamine N-methyltransferase , 2001, Proceedings of the National Academy of Sciences of the United States of America.

[14]  M. Ruiz-Echevarría,et al.  The RNA Binding Protein Pub1 Modulates the Stability of Transcripts Containing Upstream Open Reading Frames , 2000, Cell.

[15]  P. McGraw,et al.  The isolation and characterization in yeast of a gene for Arabidopsis S-adenosylmethionine:phospho-ethanolamine N-methyltransferase. , 2000, Plant physiology.

[16]  M. Mayer,et al.  A Dual Upstream Open Reading Frame-based Autoregulatory Circuit Controlling Polyamine-responsive Translation* , 2005, Journal of Biological Chemistry.

[17]  M. Bevan,et al.  GUS fusions: beta‐glucuronidase as a sensitive and versatile gene fusion marker in higher plants. , 1987, The EMBO journal.