Translational inhibition mediated by a short upstream open reading frame in the human cytomegalovirus gpUL4 (gp48) transcript.
The human cytomegalovirus (CMV) virion glycoprotein gpUL4 (gp48) gene expresses a transcript that contains three AUG codons upstream from the one used to initiate synthesis of the gp48 protein. Previously we reported that the second of these AUG codons, AUG2, was necessary but insufficient for inhibition of downstream translation (M. Schleiss, C. R. Degnin, and A. P. Geballe, J. Virol. 65:6782-6789, 1991). We now demonstrate that the coding information of the upstream open reading frame initiated by AUG2 (uORF2) is critical for the inhibitory signal. Several missense mutations, particularly those involving the carboxy-terminal codons of uORF2, inactivate the inhibitory signal, while mutations that preserve the coding content of uORF2 uniformly retain the inhibitory signal. The uORF2 termination codon is essential for inhibition, but leader sequences further downstream are not critical. Conservation of uORF2 among clinical strains of CMV suggests that uORF2 provides an important function in the CMV infectious cycle. Although these results indicate that the peptide product of uORF2 mediates the inhibitory effect, we demonstrate that the uORF2 signal acts only in cis, and we propose a model of inhibition by the gp48 uORF2 signal.
Department of Molecular Medicine, Fred Hutchinson Cancer Research Center C2-023, Seattle, Washington 98104-2092.
C R
CR
Degnin
M R
MR
Schleiss
J
J
Cao
A P
AP
Geballe
D000595:Amino Acid Sequence; D001483:Base Sequence; D002460:Cell Line; D003062:Codon / Q000235:genetics; D003587:Cytomegalovirus / Q000235:genetics*; D015967:Gene Expression Regulation, Viral; D005809:Genes, Regulator; D006801:Humans; D008969:Molecular Sequence Data; D016296:Mutagenesis; D009838:Oligodeoxyribonucleotides; D016366:Open Reading Frames; D010957:Plasmids; D011132:Polyribosomes / Q000378:metabolism; D014176:Protein Biosynthesis; D021382:Protein Sorting Signals / Q000096:biosynthesis / Q000235:genetics; D011994:Recombinant Proteins / Q000096:biosynthesis; D015183:Restriction Mapping; D017386:Sequence Homology, Amino Acid; D012689:Sequence Homology, Nucleic Acid; D013728:Terminator Regions, Genetic; D014162:Transfection; D014759:Viral Envelope Proteins / Q000096:biosynthesis*; D001616:beta-Galactosidase / Q000096:biosynthesis
D003062:Codon; D009838:Oligodeoxyribonucleotides; D021382:Protein Sorting Signals; D011994:Recombinant Proteins; C082590:UL4 protein, Human cytomegalovirus; D014759:Viral Envelope Proteins; D001616:beta-Galactosidase