Detection and characterization of degradative intermediates of avian apo very low density lipoprotein II mRNA present in estrogen-treated birds and following destabilization by hormone withdrawal.
The apo very low density lipoprotein II (apoVLDLII) gene is dormant in embryos, chicks, and roosters but can be activated by estrogen. ApoVLDLII mRNA is relatively stable in estrogen-treated birds. However, its stability decreases 4-5-fold following withdrawal of hormone. We have characterized degradative intermediates of apoVLDLII mRNA detected in liver total RNA from estrogen-treated birds and searched for alterations in the pattern of intermediates that occur upon hormone-withdrawal. Primer extension and S1 nuclease analyses have demonstrated that these intermediates consist of fragments of the molecule with intact 5' ends but which lack various 3' regions. Estrogen withdrawal results in a decrease in the steady state levels of several of these intermediates and the detection of two new species. The end points of the major fragments present in RNA from both estrogen-treated and withdrawn birds all map in, or within four nucleotides of, the tetranucleotide, GAUG. The two fragments detected only in RNA from withdrawn birds have 3' ends that immediately precede the sequence, CAGU. Based on secondary structures predicted by a global folding program, the end points also appear to be preferentially located in, or at the base of, internal "bulge-loops".
Cochrane
A
A
Department of Biochemistry, Queen's University, Kingston, Ontario, Canada.
Deeley
R G
RG
D000818:Animals; D001053:Apolipoproteins / Q000235:genetics* / Q000378:metabolism; D001483:Base Sequence; D015152:Blotting, Northern; D002645:Chickens; D004958:Estradiol / Q000494:pharmacology*; D005786:Gene Expression Regulation; D006860:Hydrogen Bonding; D008079:Lipoproteins, VLDL / Q000235:genetics* / Q000378:metabolism; D008099:Liver / Q000502:physiology; D008297:Male; D008969:Molecular Sequence Data; D009690:Nucleic Acid Conformation; D012333:RNA, Messenger / Q000378:metabolism* / Q000648:ultrastructure
D001053:Apolipoproteins; D008079:Lipoproteins, VLDL; D012333:RNA, Messenger; D004958:Estradiol