The DNA sequence coding for the 5' untranslated region of herpes simplex virus type 1 ICP22 mRNA mediates a high level of gene expression.
The sequence coding for the 5' untranslated region (UTR) of ICP22 mRNA of herpes simplex virus type 1 has been tested for its ability to regulate gene expression. This sequence was placed in frame with the chloramphenicol acetyltransferase (CAT) coding sequence and under the control of the simian virus 40 early promoter-enhancer. Under these conditions, the sequence coding for the 5'UTR led to an increase of about 13-fold in CAT activity, measured during transient expression. The use of mutants with progressive deletions within the sequence coding for the 5'UTR allowed localization of the sequence responsible for the enhancement of gene expression to the first exon of the ICP22 gene. Precise quantification of hybrid ICP22-CAT mRNA showed that the sequence coding for the 5'UTR induced an increase in the amounts of transcripts, which resulted in a parallel increase in CAT activity. This increase in the level of hybrid ICP22-CAT mRNA is not the result of an increase in mRNA stability, nor is it due to more efficient nucleo-cytoplasmic transport of the transcripts. Moreover, the distribution of hybrid mRNA in the different ribosomal populations indicates that the 5'UTR of ICP22 mRNA does not induce a preferential recruitment of the transcripts by the translational apparatus. Taken together, these results indicate that a cis-acting element located in the sequence coding for the 5'UTR of ICP22 mRNA can mediate a high level of gene expression independently of the viral promoter and of viral trans-acting factors.
Greco
A
A
Immuno-Virologie Moléculaire et Cellulaire, CNRS UMR30, Faculté de Médecine, Lyon, France.
Simonin
D
D
Diaz
J J
JJ
Barjhoux
L
L
Kindbeiter
K
K
Madjar
J J
JJ
Massé
T
T
D001483:Base Sequence; D015500:Chloramphenicol O-Acetyltransferase / Q000096:biosynthesis; D004279:DNA, Viral / Q000235:genetics; D015967:Gene Expression Regulation, Viral / Q000235:genetics*; D017781:Genes, Immediate-Early / Q000235:genetics; D006367:HeLa Cells; D018259:Herpesvirus 1, Human / Q000235:genetics*; D006801:Humans; D017874:Immediate-Early Proteins; D007438:Introns; D008969:Molecular Sequence Data; D011132:Polyribosomes / Q000378:metabolism; D012326:RNA Splicing; D012333:RNA, Messenger / Q000096:biosynthesis* / Q000235:genetics / Q000378:metabolism; D012045:Regulatory Sequences, Nucleic Acid / Q000235:genetics*; D017384:Sequence Deletion / Q000235:genetics; D014158:Transcription, Genetic; D014162:Transfection; D014407:Tumor Cells, Cultured; D014764:Viral Proteins / Q000235:genetics*; D054334:Viral Regulatory and Accessory Proteins
D004279:DNA, Viral; C057294:ICP22 protein, human herpesvirus 1; D017874:Immediate-Early Proteins; D012333:RNA, Messenger; D014764:Viral Proteins; D054334:Viral Regulatory and Accessory Proteins; C078424:EUS1 protein, Equine herpesvirus 1; D015500:Chloramphenicol O-Acetyltransferase