Molecular cloning of an amino acid-regulated mRNA (amino acid starvation-induced) in rat hepatoma cells.
Using the combination of a subtracted library and differential hybridization, a 409-base pair cDNA was identified that corresponds to a mRNA that is induced 2-3-fold when rat Fao hepatoma cells are subjected to amino acid starvation for 12 h. While this mRNA species was induced during starvation, others such as beta-actin, Cu-Zn superoxide dismutase, glyceraldehyde-3-P, and histone H4 were decreased in abundance to 25-50% of their original levels. The induction of the amino acid starvation-induced (ASI) mRNA was repressed when starved cells were returned to a medium supplemented with amino acids. Tissue distribution analysis showed the ASI mRNA, approximately 650 base pairs in length, to be present in every rat tissue tested. The cDNA clone has been sequenced and appears to correspond to the 3'-most end of the mRNA. The cDNA sequence includes the poly(A) tail, two potential polyadenylation signal sequences, and an open reading frame that we presume to be a portion of the coding sequence. The ASI cDNA will be used to investigate the molecular mechanisms for amino acid-dependent regulation of protein expression by mammalian cells.
Department of Biochemistry and Molecular Biology, J. Hillis Miller Health Center, University of Florida, College of Medicine, Gainesville 32610.
N F
NF
Shay
H S
HS
Nick
M S
MS
Kilberg
D000595:Amino Acid Sequence; D000596:Amino Acids / Q000378:metabolism* / Q000494:pharmacology; D000818:Animals; D001483:Base Sequence; D002460:Cell Line; D003001:Cloning, Molecular / Q000379:methods; D004273:DNA, Neoplasm / Q000235:genetics / Q000302:isolation & purification; D015723:Gene Library; D008114:Liver Neoplasms, Experimental; D008969:Molecular Sequence Data; D012333:RNA, Messenger / Q000096:biosynthesis / Q000235:genetics*; D012334:RNA, Neoplasm / Q000235:genetics / Q000302:isolation & purification
D000596:Amino Acids; D004273:DNA, Neoplasm; D012333:RNA, Messenger; D012334:RNA, Neoplasm