Purification of a RNA-binding protein from rat liver. Identification as ferritin L chain and determination of the RNA/protein binding characteristics - PubMed (original) (raw)
. 1997 Aug 8;272(32):20222-9.
doi: 10.1074/jbc.272.32.20222.
Affiliations
- PMID: 9242700
- DOI: 10.1074/jbc.272.32.20222
Free article
Purification of a RNA-binding protein from rat liver. Identification as ferritin L chain and determination of the RNA/protein binding characteristics
T Heise et al. J Biol Chem. 1997.
Free article
Abstract
In cultured rat hepatocytes the degradation of phosphoenolpyruvate carboxykinase mRNA might be regulated by protein(s), which by binding to the mRNA alter its stability. The 3'-untranslated region of phosphoenolpyruvate carboxykinase mRNA as a potential target was used to select RNA-binding protein(s) from rat liver by the use of gel retardation assays. A cytosolic protein was isolated, which bound to the phosphoenolpyruvate carboxykinase mRNA 3'-untranslated region and other in vitro synthesized RNAs. The protein was purified to homogeneity; it had an apparent molecular mass of 400 kDa and consisted of identical subunits with an apparent size of 24.5 kDa. Sequence analysis of a tryptic peptide from the 24.5-kDa protein revealed its identity with rat ferritin light chain. Binding of ferritin to RNA was abolished after phosphorylation with cAMP-dependent protein kinase and was augmented after dephosphorylation with alkaline phosphatase. Weak binding was observed in extracts from okadaic acid-treated cultured hepatocytes compared with untreated cells. Preincubation of ferritin with an anti-phosphoserine or an anti-phosphothreonine antibody attenuated binding to RNA, while an anti-phosphotyrosine antibody generated a supershift indicating that phosphoserine and phosphothreonine but not phosphotyrosine residues were in close proximity to the RNA-binding region. Ferritin is the iron storage protein in the liver. Binding of ferritin to RNA was diminished in the presence of increasing iron concentrations, whereas the iron chelator desferal was without effect. It is concluded that ferritin might function as RNA-binding protein and that it may have important functions in the general regulation of cellular RNA metabolism.
Similar articles
- A cAMP-regulated RNA-binding protein that interacts with phosphoenolpyruvate carboxykinase (GTP) mRNA.
Nachaliel N, Jain D, Hod Y. Nachaliel N, et al. J Biol Chem. 1993 Nov 15;268(32):24203-9. J Biol Chem. 1993. PMID: 8226967 - Parallel acceleration of phosphoenolpyruvate carboxykinase mRNA degradation and increase in ribonuclease activity induced by insulin in cultured rat hepatocytes.
Heise T, Krones A, Nath A, Jungermann K, Christ B. Heise T, et al. Biol Chem. 1998 Jul;379(7):875-83. doi: 10.1515/bchm.1998.379.7.875. Biol Chem. 1998. PMID: 9705151 - Isolation, characterization, and functional studies of rat liver iron regulatory protein 1.
Eisenstein RS, Barton HA, Pettingell WH Jr, Bomford AB. Eisenstein RS, et al. Arch Biochem Biophys. 1997 Jul 1;343(1):81-91. doi: 10.1006/abbi.1997.0144. Arch Biochem Biophys. 1997. PMID: 9210649 - Translational regulation of ferritin synthesis by iron.
Eisenstein RS, Munro HN. Eisenstein RS, et al. Enzyme. 1990;44(1-4):42-58. doi: 10.1159/000468746. Enzyme. 1990. PMID: 2133657 Review. - The ferritins: molecular properties, iron storage function and cellular regulation.
Harrison PM, Arosio P. Harrison PM, et al. Biochim Biophys Acta. 1996 Jul 31;1275(3):161-203. doi: 10.1016/0005-2728(96)00022-9. Biochim Biophys Acta. 1996. PMID: 8695634 Review.
Cited by
- Characterization of nuclear RNases that cleave hepatitis B virus RNA near the La protein binding site.
Heise T, Guidotti LG, Chisari FV. Heise T, et al. J Virol. 2001 Aug;75(15):6874-83. doi: 10.1128/JVI.75.15.6874-6883.2001. J Virol. 2001. PMID: 11435567 Free PMC article. - Hepatitis B virus RNA-binding proteins associated with cytokine-induced clearance of viral RNA from the liver of transgenic mice.
Heise T, Guidotti LG, Cavanaugh VJ, Chisari FV. Heise T, et al. J Virol. 1999 Jan;73(1):474-81. doi: 10.1128/JVI.73.1.474-481.1999. J Virol. 1999. PMID: 9847353 Free PMC article.