Sequences in the rev-responsive element responsible for premature translational arrest in the human-immunodeficiency-virus-type-1 envelope.
Cell-free translation in the presence of pancreatic microsomal membranes of the full-length envelope transcript of the human immunodeficiency virus type 1 (HIV-1) yielded the expected extensively glycosylated and immunologically reactive gp160 envelope-protein precursor. In addition to this gp160, a shorter glycoprotein, which we designated gp120*, was produced due to a premature translation arrest. Utilizing kinetic experiments, pulse-chase analyses and various gp160 envelope RNA mutants, we demonstrated that the in-vitro-produced gp120* was not formed by cleavage of the gp160 precursor or by internal initiation of translation. A gp120 produced before gp160 synthesis was completed, and, independent of the gp160 proteolytic processing, has been shown to be produced and sequestered in the endoplasmic reticulum of HIV-1-infected cells [Willey, R. L., Klimkait, T., Frucht, D. M., Bonifacino, J. S. & Martin, M. A. (1991) Virology 184, 319-329]. The specific translational arrest shown to occur in vitro was found to be dependent on the Rev-responsive element, since deletion of this highly structured sequence abolished the production of gp120*. We found that the combination of two contiguous putative stem loops of the Rev-responsive element, located at nucleotides 7494-7522 and 7525-7550 of the HIV-1 Rev-responsive-element sequence, was responsible for the production of this truncated protein. To our knowledge, these stem-loop structures, distinct from that known to bind the Rev protein, represent the first example responsible for the production of alternative products by premature translational arrest in higher eukaryotes.
Ellerbrok
H
H
Oncologie Cellulaire et Moléculaire INSERM U363, Institut Cochin de Génétique Moléculaire, Paris, France.
Serpente
N
N
Pancino
G
G
Vanhée
C
C
D'Auriol
L
L
Sitbon
M
M
Vaquero
C
C
D001483:Base Sequence; D004279:DNA, Viral / Q000737:chemistry / Q000235:genetics; D015686:Gene Products, env / Q000096:biosynthesis / Q000235:genetics*; D015694:Gene Products, rev / Q000235:genetics*; D015752:Genes, env; D015699:HIV Envelope Protein gp120 / Q000096:biosynthesis / Q000235:genetics; D019302:HIV Envelope Protein gp160; D015497:HIV-1 / Q000235:genetics* / Q000378:metabolism; D008969:Molecular Sequence Data; D009690:Nucleic Acid Conformation; D014176:Protein Biosynthesis; D011498:Protein Precursors / Q000096:biosynthesis / Q000235:genetics* / Q000378:metabolism; D012367:RNA, Viral; D012045:Regulatory Sequences, Nucleic Acid; D013728:Terminator Regions, Genetic; D014158:Transcription, Genetic; D054321:rev Gene Products, Human Immunodeficiency Virus
D004279:DNA, Viral; D015686:Gene Products, env; D015694:Gene Products, rev; D015699:HIV Envelope Protein gp120; D019302:HIV Envelope Protein gp160; D011498:Protein Precursors; D012367:RNA, Viral; D054321:rev Gene Products, Human Immunodeficiency Virus