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Plant coilin: structural characteristics and RNA-binding properties.

Makarov V, Rakitina D, Protopopova A, Yaminsky I, Arutiunian A, Love AJ, Taliansky M, Kalinina N - PLoS ONE (2013)

Bottom Line: Atcoilin is able to bind RNA effectively and in a non-specific manner.The interaction with RNA and subsequent multimerization may facilitate coilin's function as a scaffolding protein.A model of the N-terminal domain is also proposed.

View Article: PubMed Central - PubMed

Affiliation: A. N. Belozersky Institute of Physico-Chemical Biology, Moscow State University, Moscow, Russia.

ABSTRACT
Cajal bodies (CBs) are dynamic subnuclear compartments involved in the biogenesis of ribonucleoproteins. Coilin is a major structural scaffolding protein necessary for CB formation, composition and activity. The predicted secondary structure of Arabidopsis thaliana coilin (Atcoilin) suggests that the protein is composed of three main domains. Analysis of the physical properties of deletion mutants indicates that Atcoilin might consist of an N-terminal globular domain, a central highly disordered domain and a C-terminal domain containing a presumable Tudor-like structure adjacent to a disordered C terminus. Despite the low homology in amino acid sequences, a similar type of domain organization is likely shared by human and animal coilin proteins and coilin-like proteins of various plant species. Atcoilin is able to bind RNA effectively and in a non-specific manner. This activity is provided by three RNA-binding sites: two sets of basic amino acids in the N-terminal domain and one set in the central domain. Interaction with RNA induces the multimerization of the Atcoilin molecule, a consequence of the structural alterations in the N-terminal domain. The interaction with RNA and subsequent multimerization may facilitate coilin's function as a scaffolding protein. A model of the N-terminal domain is also proposed.

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Analysis of the RNA-binding activity of Atcoilin and its mutants.The percentage of RNA complexed with the protein is plotted versus the protein concentration in the sample (µM). (A) Coilin binding to U1 and U2 snRNAs and pGEM7 RNA. (B) and (C) Binding of U1 snRNA by NOD, CTD, IDD and their mutants. Mutants compared on one plot are indicated.
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pone-0053571-g004: Analysis of the RNA-binding activity of Atcoilin and its mutants.The percentage of RNA complexed with the protein is plotted versus the protein concentration in the sample (µM). (A) Coilin binding to U1 and U2 snRNAs and pGEM7 RNA. (B) and (C) Binding of U1 snRNA by NOD, CTD, IDD and their mutants. Mutants compared on one plot are indicated.

Mentions: In this report we have tested the ability of coilin to directly interact with two snRNAs - U2 snRNA and U1 snRNA and a non-specific artificial RNA (160 nt). In the electrophoretic mobility shift assay (EMSA) the recombinant Atcoilin is able to bind all these substrates with similar characteristics: apparent Kd values are 0.18–0.2 µM, with 50% of the RNA being incorporated into the complex at the protein:RNA molar ratio of 2∶1, and 100% of the RNA was bound at the molar ratio of 6∶1 (Figure 3B, 4A). The Hill constant is about 1.2±0.1, and is indicative of low cooperative binding.


Plant coilin: structural characteristics and RNA-binding properties.

Makarov V, Rakitina D, Protopopova A, Yaminsky I, Arutiunian A, Love AJ, Taliansky M, Kalinina N - PLoS ONE (2013)

Analysis of the RNA-binding activity of Atcoilin and its mutants.The percentage of RNA complexed with the protein is plotted versus the protein concentration in the sample (µM). (A) Coilin binding to U1 and U2 snRNAs and pGEM7 RNA. (B) and (C) Binding of U1 snRNA by NOD, CTD, IDD and their mutants. Mutants compared on one plot are indicated.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC3539977&req=5

pone-0053571-g004: Analysis of the RNA-binding activity of Atcoilin and its mutants.The percentage of RNA complexed with the protein is plotted versus the protein concentration in the sample (µM). (A) Coilin binding to U1 and U2 snRNAs and pGEM7 RNA. (B) and (C) Binding of U1 snRNA by NOD, CTD, IDD and their mutants. Mutants compared on one plot are indicated.
Mentions: In this report we have tested the ability of coilin to directly interact with two snRNAs - U2 snRNA and U1 snRNA and a non-specific artificial RNA (160 nt). In the electrophoretic mobility shift assay (EMSA) the recombinant Atcoilin is able to bind all these substrates with similar characteristics: apparent Kd values are 0.18–0.2 µM, with 50% of the RNA being incorporated into the complex at the protein:RNA molar ratio of 2∶1, and 100% of the RNA was bound at the molar ratio of 6∶1 (Figure 3B, 4A). The Hill constant is about 1.2±0.1, and is indicative of low cooperative binding.

Bottom Line: Atcoilin is able to bind RNA effectively and in a non-specific manner.The interaction with RNA and subsequent multimerization may facilitate coilin's function as a scaffolding protein.A model of the N-terminal domain is also proposed.

View Article: PubMed Central - PubMed

Affiliation: A. N. Belozersky Institute of Physico-Chemical Biology, Moscow State University, Moscow, Russia.

ABSTRACT
Cajal bodies (CBs) are dynamic subnuclear compartments involved in the biogenesis of ribonucleoproteins. Coilin is a major structural scaffolding protein necessary for CB formation, composition and activity. The predicted secondary structure of Arabidopsis thaliana coilin (Atcoilin) suggests that the protein is composed of three main domains. Analysis of the physical properties of deletion mutants indicates that Atcoilin might consist of an N-terminal globular domain, a central highly disordered domain and a C-terminal domain containing a presumable Tudor-like structure adjacent to a disordered C terminus. Despite the low homology in amino acid sequences, a similar type of domain organization is likely shared by human and animal coilin proteins and coilin-like proteins of various plant species. Atcoilin is able to bind RNA effectively and in a non-specific manner. This activity is provided by three RNA-binding sites: two sets of basic amino acids in the N-terminal domain and one set in the central domain. Interaction with RNA induces the multimerization of the Atcoilin molecule, a consequence of the structural alterations in the N-terminal domain. The interaction with RNA and subsequent multimerization may facilitate coilin's function as a scaffolding protein. A model of the N-terminal domain is also proposed.

Show MeSH