Purification by affinity chromatography of H1 RNA-Binding Proteins from rat brain

Scaturro, M; Sala, A; Cutrona, G; Raimondi, L; Cannino, G; Pucci Minafra, I

Risultato della ricerca: Article

6 Citazioni (Scopus)

Abstract

Post-transcriptional regulation of mRNA metabolism is involved in processes as different as cell fate specification in development and cell response to a large variety of environmental cues. Regulation of all steps of RNA metabolism depends on RNA-binding proteins (RBPs). By using a T1 RNase protection assay, we previously identified three H1° RNA-binding factors (p40, p70 and p110), highly expressed in the rat brain. Here we report enrichment of these factors from brain extracts, obtained by affinity chromatography of biotinylated H1° RNA-protein complexes on streptavidin-conjugated paramagnetic particles. The purified proteins maintain RNA-binding ability and preference for histone messages. Thanks to the efforts of a number of laboratories in the last decade, it is now clear that a central component in the control of gene expression involves post-transcriptional regulation of mRNA metabolism (reviewed in refs. 1-7). Post-transcriptional regulation of time and site in which a given mRNA must be translated and its duration depend on two main classes of RNA-binding proteins: i) RNA chaperones, that assist folding of RNA molecules (8), and ii) tertiary-structure-binding regulatory proteins, that recognize and stabilize specific tertiary structures (9). The two classes of proteins include both nonspecific and specific RNA-binding functions (10-13). In vivo, most mRNAs are actually present as large ribonucleoprotein complexes, the assembly of which probably requires a series of events including formation of RNA secondary structures and selective stabilization of specific tertiary structures by proteins (9,14-15). In the last few years we have been looking for proteins able to bind mRNAs encoding the histone variants H1° and H3.3 (16-19). In searching for such factors, we adopted and/or developed a series of different experimental approaches. In particular, by using a T1 RNase-protection assay, we previously identified three H1° RNA-binding factors (p40, p70 and p110), highly expressed in the rat brain (17). Herein we describe purification of H1° RNA-binding factors from rat brain by an affinity chromatography protocol that made use of an in vitro-transcribed, biotinylated H1° RNA and of streptavidin-conjugated paramagnetic particles.
Lingua originaleEnglish
pagine (da-a)509-513
Numero di pagine5
RivistaInternational Journal of Molecular Medicine
Volume11
Stato di pubblicazionePublished - 2003

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RNA-Binding Proteins
Affinity Chromatography
RNA
Brain
Messenger RNA
Histones
Ribonuclease T1
Streptavidin
RNA Folding
Proteins
Aptitude
Ribonucleoproteins
Tertiary Protein Structure
Cues
Carrier Proteins
Gene Expression

All Science Journal Classification (ASJC) codes

  • Genetics

Cita questo

Scaturro, M; Sala, A; Cutrona, G; Raimondi, L; Cannino, G; Pucci Minafra, I (2003). Purification by affinity chromatography of H1 RNA-Binding Proteins from rat brain. International Journal of Molecular Medicine, 11, 509-513.

Purification by affinity chromatography of H1 RNA-Binding Proteins from rat brain. / Scaturro, M; Sala, A; Cutrona, G; Raimondi, L; Cannino, G; Pucci Minafra, I.

In: International Journal of Molecular Medicine, Vol. 11, 2003, pag. 509-513.

Risultato della ricerca: Article

Scaturro, M; Sala, A; Cutrona, G; Raimondi, L; Cannino, G; Pucci Minafra, I 2003, 'Purification by affinity chromatography of H1 RNA-Binding Proteins from rat brain', International Journal of Molecular Medicine, vol. 11, pagg. 509-513.
Scaturro, M; Sala, A; Cutrona, G; Raimondi, L; Cannino, G; Pucci Minafra, I. Purification by affinity chromatography of H1 RNA-Binding Proteins from rat brain. International Journal of Molecular Medicine. 2003;11:509-513.
Scaturro, M; Sala, A; Cutrona, G; Raimondi, L; Cannino, G; Pucci Minafra, I. / Purification by affinity chromatography of H1 RNA-Binding Proteins from rat brain. In: International Journal of Molecular Medicine. 2003 ; Vol. 11. pagg. 509-513.
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abstract = "Post-transcriptional regulation of mRNA metabolism is involved in processes as different as cell fate specification in development and cell response to a large variety of environmental cues. Regulation of all steps of RNA metabolism depends on RNA-binding proteins (RBPs). By using a T1 RNase protection assay, we previously identified three H1° RNA-binding factors (p40, p70 and p110), highly expressed in the rat brain. Here we report enrichment of these factors from brain extracts, obtained by affinity chromatography of biotinylated H1° RNA-protein complexes on streptavidin-conjugated paramagnetic particles. The purified proteins maintain RNA-binding ability and preference for histone messages. Thanks to the efforts of a number of laboratories in the last decade, it is now clear that a central component in the control of gene expression involves post-transcriptional regulation of mRNA metabolism (reviewed in refs. 1-7). Post-transcriptional regulation of time and site in which a given mRNA must be translated and its duration depend on two main classes of RNA-binding proteins: i) RNA chaperones, that assist folding of RNA molecules (8), and ii) tertiary-structure-binding regulatory proteins, that recognize and stabilize specific tertiary structures (9). The two classes of proteins include both nonspecific and specific RNA-binding functions (10-13). In vivo, most mRNAs are actually present as large ribonucleoprotein complexes, the assembly of which probably requires a series of events including formation of RNA secondary structures and selective stabilization of specific tertiary structures by proteins (9,14-15). In the last few years we have been looking for proteins able to bind mRNAs encoding the histone variants H1° and H3.3 (16-19). In searching for such factors, we adopted and/or developed a series of different experimental approaches. In particular, by using a T1 RNase-protection assay, we previously identified three H1° RNA-binding factors (p40, p70 and p110), highly expressed in the rat brain (17). Herein we describe purification of H1° RNA-binding factors from rat brain by an affinity chromatography protocol that made use of an in vitro-transcribed, biotinylated H1° RNA and of streptavidin-conjugated paramagnetic particles.",
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AB - Post-transcriptional regulation of mRNA metabolism is involved in processes as different as cell fate specification in development and cell response to a large variety of environmental cues. Regulation of all steps of RNA metabolism depends on RNA-binding proteins (RBPs). By using a T1 RNase protection assay, we previously identified three H1° RNA-binding factors (p40, p70 and p110), highly expressed in the rat brain. Here we report enrichment of these factors from brain extracts, obtained by affinity chromatography of biotinylated H1° RNA-protein complexes on streptavidin-conjugated paramagnetic particles. The purified proteins maintain RNA-binding ability and preference for histone messages. Thanks to the efforts of a number of laboratories in the last decade, it is now clear that a central component in the control of gene expression involves post-transcriptional regulation of mRNA metabolism (reviewed in refs. 1-7). Post-transcriptional regulation of time and site in which a given mRNA must be translated and its duration depend on two main classes of RNA-binding proteins: i) RNA chaperones, that assist folding of RNA molecules (8), and ii) tertiary-structure-binding regulatory proteins, that recognize and stabilize specific tertiary structures (9). The two classes of proteins include both nonspecific and specific RNA-binding functions (10-13). In vivo, most mRNAs are actually present as large ribonucleoprotein complexes, the assembly of which probably requires a series of events including formation of RNA secondary structures and selective stabilization of specific tertiary structures by proteins (9,14-15). In the last few years we have been looking for proteins able to bind mRNAs encoding the histone variants H1° and H3.3 (16-19). In searching for such factors, we adopted and/or developed a series of different experimental approaches. In particular, by using a T1 RNase-protection assay, we previously identified three H1° RNA-binding factors (p40, p70 and p110), highly expressed in the rat brain (17). Herein we describe purification of H1° RNA-binding factors from rat brain by an affinity chromatography protocol that made use of an in vitro-transcribed, biotinylated H1° RNA and of streptavidin-conjugated paramagnetic particles.

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