PMID: 3762527Jan 1, 1986Paper

In vitro release of alpha 1-acid glycoprotein RNA sequences shows fidelity with the acute phase response in vivo

Molecular Biology Reports
G A ClawsonE A Smuckler

Abstract

The acute phase reaction of rat liver to subcutaneous turpentine challenge results in a 20- to 100-fold increase in alpha 1-acid glycoprotein (alpha AGP) mRNA. We utilized this response to establish conditions appropriate for study of RNA transport in vitro using hybridization with 32P-labeled exon and intron alpha AGP sequences. Contamination of nuclear preparations by membrane-absorbed cytoplasmic RNA was eliminated by detergent-rinsing. The in vitro incubation conditions that most reflected the in vivo state required RNase inhibitor (purified from placenta), polyvinylpyrrolidone to prevent nuclear swelling, and addition of ATP. Under these circumstances, alpha AGP sequences were transported only from turpentine-stimulated preparations, were found only in poly(A)+ RNA, and were the same size as authentic cytoplasmic mRNA. Omission of polyvinylpyrrolidone resulted in release of some alpha AGP sequences in smaller, more heterogeneous poly(A)- RNA, and leakage of some alpha AGP sequences was observed from control preparations. Omission of ATP resulted in restriction of mature alpha AGP mRNA to the nucleus. In contrast to alpha AGP mRNA, transport of albumin mRNA was decreased 3-4X in turpentine-treated preparations. The largest ...Continue Reading

References

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Citations

Jul 1, 1985·Molecular and Cellular Biochemistry·G A ClawsonE A Smuckler
Dec 16, 1991·Clinica Chimica Acta; International Journal of Clinical Chemistry·C B EapP Baumann
Nov 4, 2000·Biochimica Et Biophysica Acta·T FournierD Porquet
May 13, 1999·Nutrition·Y Ingenbleek, L Bernstein
Dec 1, 1996·Academic Emergency Medicine : Official Journal of the Society for Academic Emergency Medicine·R GoldbergM L McConkie
Jul 31, 1991·Biochemical and Biophysical Research Communications·B K Ray, A Ray
May 29, 1992·Biochemical and Biophysical Research Communications·B K Ray, A Ray

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