Initiation of translation by non-AUG codons in human T-cell lymphotropic virus type I mRNA encoding both Rex and Tax regulatory proteins.
Human T-cell lymphotropic virus type I (HTLV-I) double-spliced mRNA exhibits two GUG and two CUG codons upstream to, and in frame with, the sequences encoding Rex and Tax regulatory proteins, respectively. To verify whether these GUG and CUG codons could be used as additional initiation codons of translation, two chimeric constructs were built for directing the synthesis of either Rex-CAT or Tax-CAT fusion proteins. In both cases, the CAT reporter sequence was inserted after the Tax AUG codon and in frame with either the Rex or Tax AUG codon. Under transient expression of these constructs, other proteins of higher molecular mass were synthesized in addition to the expected Rex-CAT and Tax-CAT proteins. The potential non-AUG initiation codons were exchanged for either an AUG codon or a non-initiation codon. This allowed us to demonstrate that the two GUG codons in frame with the Rex coding sequence, and only the second CUG in frame with the Tax coding sequence, were used as additional initiation codons. In HTLV-I infected cells, two Rex and one Tax additional proteins were detected that exhibited molecular mass compatible with the use of the two GUG and the second CUG as additional initiation codons of translation. Comparison of the HTLV-I proviral DNA sequence with that of other HTLV-related retroviruses revealed a striking conservation of the three non-AUG initiation codons, strongly suggesting their use for the synthesis of additional Rex and Tax proteins.
Corcelette
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Laboratoire de Biologie Moléculaire et Cellulaire, INSERM U 369, Faculté de Médecine Lyon-RTH Laennec, 7 rue Guillaume Paradin, 69372 Lyon Cedex 08, France.
Massé
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Madjar
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D001483:Base Sequence; D015500:Chloramphenicol O-Acetyltransferase / Q000096:biosynthesis / Q000235:genetics; D018387:Codon, Initiator / Q000235:genetics; D017931:DNA Primers / Q000235:genetics; D016370:Gene Products, rex / Q000096:biosynthesis / Q000235:genetics; D016356:Gene Products, tax / Q000096:biosynthesis / Q000235:genetics; D005814:Genes, Viral; D005822:Genetic Vectors; D006367:HeLa Cells; D015368:Human T-lymphotropic virus 1 / Q000235:genetics* / Q000378:metabolism; D006801:Humans; D001909:Leukemia Virus, Bovine / Q000235:genetics; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D010442:Peptide Chain Initiation, Translational; D012333:RNA, Messenger / Q000235:genetics*; D012367:RNA, Viral / Q000235:genetics*; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000235:genetics; D012689:Sequence Homology, Nucleic Acid; D015303:Simian T-lymphotropic virus 1 / Q000235:genetics; D013045:Species Specificity; D014162:Transfection
D018387:Codon, Initiator; D017931:DNA Primers; D016370:Gene Products, rex; D016356:Gene Products, tax; D012333:RNA, Messenger; D012367:RNA, Viral; D011993:Recombinant Fusion Proteins; D015500:Chloramphenicol O-Acetyltransferase