Isolation and Characterization of a Folate Receptor mRNA-binding trans-Factor from Human Placenta. Evidence favoring identity with heterogeneous nuclear ribonucleoprotein E1

Xiangli Xiao, Ying Sheng Tang, Janet Y. Mackins, Xin Lai Sun, Hiremagalur N. Jayaram, Deborah K. Hansen, Asok Antony

Research output: Contribution to journalArticle

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Abstract

The interaction of an 18-base cis-element in the 5′-untranslated region of human folate receptor (FR)-α mRNA with a cytosolic trans-factor protein is critical for the translation of FR (Sun, X.-L., and Antony, A. C. (1996) J. Biol. Chem. 271, 25539-25547). This trans-factor was isolated to apparent homogeneity as a 43- and 38-kDa doublet from human placenta using poly(U)-Sepharose, followed by preparative sodium dodecyl sulfate-polyacrylamide gel electrophoresis and electro-elution as major purification steps. Amino acid microsequencing of two cyanogen bromide-generated peptide fragments of the 43-kDa trans-factor revealed complete identity with 43-kDa heterogeneous nuclear ribonucleoprotein E1 (hnRNP E1). Purified specific rabbit anti-hnRNP E1 peptide antibodies (generated against a synthetic oligopeptide that was not represented in microsequenced peptides of the trans-factor) also recognized the purified trans-factor on Western blots. Conversely, the 18-base FR RNA cis-element also bound hnRNP E1 protein on Northwestern blots. Moreover, a 19-base RNA cis-element in the 3′-untranslated region of 15-lipoxygenase mRNA that is known to bind hnRNP E1 also interacted with placental 43-kDa trans-factor. In addition, several murine tissues containing a hnRNP E1-related protein (also known as αCP-1) readily interacted with the 18-base FR RNA cis-element. Finally, anti-hnRNP E1 anti-bodies specifically inhibited translation of FR in vitro in a dose-dependent manner, and the antibody effect could be reversed in a dose-dependent manner by either purified trans-factor or hnRNP E1. Collectively, the data favor identity of the FR mRNA-binding trans-factor and hnRNP E1, confirm its critical role in the translation of FR, and highlight yet another role of multifunctional hnRNP E1 in eukaryotic mRNA regulation.

Original languageEnglish
Pages (from-to)41510-41517
Number of pages8
JournalJournal of Biological Chemistry
Volume276
Issue number44
DOIs
StatePublished - Nov 2 2001

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Heterogeneous-Nuclear Ribonucleoproteins
Folic Acid
Placenta
Messenger RNA
RNA
Arachidonate 15-Lipoxygenase
Cyanogen Bromide
Oligopeptides
Peptides
Peptide Fragments
Proteins
Antibodies
5' Untranslated Regions
3' Untranslated Regions
Solar System
Electrophoresis
Sodium Dodecyl Sulfate
Sun
Purification
Polyacrylamide Gel Electrophoresis

ASJC Scopus subject areas

  • Biochemistry

Cite this

Isolation and Characterization of a Folate Receptor mRNA-binding trans-Factor from Human Placenta. Evidence favoring identity with heterogeneous nuclear ribonucleoprotein E1. / Xiao, Xiangli; Tang, Ying Sheng; Mackins, Janet Y.; Sun, Xin Lai; Jayaram, Hiremagalur N.; Hansen, Deborah K.; Antony, Asok.

In: Journal of Biological Chemistry, Vol. 276, No. 44, 02.11.2001, p. 41510-41517.

Research output: Contribution to journalArticle

Xiao, Xiangli ; Tang, Ying Sheng ; Mackins, Janet Y. ; Sun, Xin Lai ; Jayaram, Hiremagalur N. ; Hansen, Deborah K. ; Antony, Asok. / Isolation and Characterization of a Folate Receptor mRNA-binding trans-Factor from Human Placenta. Evidence favoring identity with heterogeneous nuclear ribonucleoprotein E1. In: Journal of Biological Chemistry. 2001 ; Vol. 276, No. 44. pp. 41510-41517.
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abstract = "The interaction of an 18-base cis-element in the 5′-untranslated region of human folate receptor (FR)-α mRNA with a cytosolic trans-factor protein is critical for the translation of FR (Sun, X.-L., and Antony, A. C. (1996) J. Biol. Chem. 271, 25539-25547). This trans-factor was isolated to apparent homogeneity as a 43- and 38-kDa doublet from human placenta using poly(U)-Sepharose, followed by preparative sodium dodecyl sulfate-polyacrylamide gel electrophoresis and electro-elution as major purification steps. Amino acid microsequencing of two cyanogen bromide-generated peptide fragments of the 43-kDa trans-factor revealed complete identity with 43-kDa heterogeneous nuclear ribonucleoprotein E1 (hnRNP E1). Purified specific rabbit anti-hnRNP E1 peptide antibodies (generated against a synthetic oligopeptide that was not represented in microsequenced peptides of the trans-factor) also recognized the purified trans-factor on Western blots. Conversely, the 18-base FR RNA cis-element also bound hnRNP E1 protein on Northwestern blots. Moreover, a 19-base RNA cis-element in the 3′-untranslated region of 15-lipoxygenase mRNA that is known to bind hnRNP E1 also interacted with placental 43-kDa trans-factor. In addition, several murine tissues containing a hnRNP E1-related protein (also known as αCP-1) readily interacted with the 18-base FR RNA cis-element. Finally, anti-hnRNP E1 anti-bodies specifically inhibited translation of FR in vitro in a dose-dependent manner, and the antibody effect could be reversed in a dose-dependent manner by either purified trans-factor or hnRNP E1. Collectively, the data favor identity of the FR mRNA-binding trans-factor and hnRNP E1, confirm its critical role in the translation of FR, and highlight yet another role of multifunctional hnRNP E1 in eukaryotic mRNA regulation.",
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AU - Antony, Asok

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