Interaction of the eucaryotic peptide chain initiation factor eIF-4A with the specific elements at the 5'-untranslated sequence of human asparagine synthetase mRNA.
The resistance of certain tumor cells to the chemotherapeutic agent L-asparaginase has often been found to be associated with the presence of asparagine synthetase activity. In an attempt to study the translational regulation of the asparagine synthetase gene, the 5'-untranslated region of human asparagine synthetase cDNA was mapped by antisense oligonucleotide-mediated hybrid arrest translation in reticulocyte lysate. Three consecutive cis-acting regulatory elements, spanning from -60 to -120 bases from the initiation codon, in the 5'-untranslated region of the asparagine synthetase gene, were identified. T1 RNase footprinting analysis showed that those regulatory elements can be protected from T1 digestion when incubated with reticulocyte lysate. A 46-kDa trans-acting protein factor that interacts with the cis-acting regulatory element of asparagine synthetase mRNA was detected. This 46-kDa protein factor is most likely to be the eucaryotic peptide chain initiation factor eIF-4A as determined by immunoprecipitation experiments using a monoclonal antibody raised against reticulocyte eIF-4A.
Chakrabarti
R
R
Department of Biochemistry and Molecular Biology, University of Florida, Gainesville 32610.
Chakrabarti
D
D
Souba
W W
WW
Schuster
S M
SM
D001217:Aspartate-Ammonia Ligase / Q000235:genetics*; D001483:Base Sequence; D001665:Binding Sites; D039601:Eukaryotic Initiation Factor-4A; D006801:Humans; D008969:Molecular Sequence Data; D009838:Oligodeoxyribonucleotides; D010448:Peptide Initiation Factors / Q000378:metabolism*; D010957:Plasmids; D014176:Protein Biosynthesis; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism*; D012045:Regulatory Sequences, Nucleic Acid; D015183:Restriction Mapping; D006163:Ribonuclease T1; D014158:Transcription, Genetic
D009838:Oligodeoxyribonucleotides; D010448:Peptide Initiation Factors; D012333:RNA, Messenger; D039601:Eukaryotic Initiation Factor-4A; D006163:Ribonuclease T1; D001217:Aspartate-Ammonia Ligase