RNA is an integral component of chromatin that contributes to its structural organization

Chromatin structure is influenced by multiples factors, such as pH, temperature, nature and concentration of counterions, post-translational modifications of histones and binding of structural non-histone proteins. RNA is also known to contribute to the regulation of chromatin structure as chromatin...

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Veröffentlicht in:PloS one 2007-11, Vol.2 (11), p.e1182
Hauptverfasser: Rodríguez-Campos, Antonio, Azorín, Fernando
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Azorín, Fernando
description Chromatin structure is influenced by multiples factors, such as pH, temperature, nature and concentration of counterions, post-translational modifications of histones and binding of structural non-histone proteins. RNA is also known to contribute to the regulation of chromatin structure as chromatin-induced gene silencing was shown to depend on the RNAi machinery in S. pombe, plants and Drosophila. Moreover, both in Drosophila and mammals, dosage compensation requires the contribution of specific non-coding RNAs. However, whether RNA itself plays a direct structural role in chromatin is not known. Here, we report results that indicate a general structural role for RNA in eukaryotic chromatin. RNA is found associated to purified chromatin prepared from chicken liver, or cultured Drosophila S2 cells, and treatment with RNase A alters the structural properties of chromatin. Our results indicate that chromatin-associated RNAs, which account for 2%-5% of total chromatin-associated nucleic acids, are polyA(-) and show a size similar to that of the DNA contained in the corresponding chromatin fragments. Chromatin-associated RNA(s) are not likely to correspond to nascent transcripts as they are also found bound to chromatin when cells are treated with alpha-amanitin. After treatment with RNase A, chromatin fragments of molecular weight >3.000 bp of DNA showed reduced sedimentation through sucrose gradients and increased sensitivity to micrococcal nuclease digestion. This structural transition, which is observed both at euchromatic and heterochromatic regions, proceeds without loss of histone H1 or any significant change in core-histone composition and integrity.
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RNA is also known to contribute to the regulation of chromatin structure as chromatin-induced gene silencing was shown to depend on the RNAi machinery in S. pombe, plants and Drosophila. Moreover, both in Drosophila and mammals, dosage compensation requires the contribution of specific non-coding RNAs. However, whether RNA itself plays a direct structural role in chromatin is not known. Here, we report results that indicate a general structural role for RNA in eukaryotic chromatin. RNA is found associated to purified chromatin prepared from chicken liver, or cultured Drosophila S2 cells, and treatment with RNase A alters the structural properties of chromatin. Our results indicate that chromatin-associated RNAs, which account for 2%-5% of total chromatin-associated nucleic acids, are polyA(-) and show a size similar to that of the DNA contained in the corresponding chromatin fragments. 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subjects Alpha-Amanitin - chemistry
Amanitin
Animals
Biosynthesis
Chickens
Chromatin
Chromatin - chemistry
Core loss
Deoxyribonucleic acid
DNA
DNA binding proteins
Dosage compensation
Drosophila
Drosophila melanogaster
Fragments
Gene expression
Gene silencing
Genes
Genetic engineering
Genomics
Histone H1
Histones
Insects
Liver
Liver - chemistry
Medical research
Molecular biology
Molecular Biology/Chromatin Structure
Molecular weight
Non-coding RNA
Nuclease
Nucleases
Nucleic acids
Osmolar Concentration
pH effects
Post-translation
Protein binding
Proteins
Ribonuclease
Ribonuclease, Pancreatic - chemistry
Ribonucleic acid
RNA
RNA - chemistry
RNA Interference
RNA polymerase
RNA-mediated interference
Sedimentation
Sucrose
Sugar
Weight reduction
title RNA is an integral component of chromatin that contributes to its structural organization
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