Proteins of chromatin in template restriction. I. RNA synthesis in vitro

T. C. Spelsberg; L. S. Hnilica

doi:10.1016/0005-2787(71)90560-0

Proteins of chromatin in template restriction. I. RNA synthesis in vitro

T. C. Spelsberg, L. S. Hnilica

Biochemistry and Molecular Biology

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Abstract

1. 1. Selective extraction of the very lysine-rich histone fraction F1 does not increase the activity of chromatin as a template for RNA synthesis in vitro. Removal of either arginine-rich histones F2a and F3 or lysine-rich histones F2b (together with F1) results in an increase of the template activity of chromatin. 2. 2. (NH₄)₂SO₄ removes selectively the lysine-rich histone fractions F1 and F2b. Even in 1.0 M (NH₄)₂SO₄ the arginine-rich histones F2a and F3 remain associated with chromatin. 3. 3. (NH₄)₂SO₄ in 0.2 M concentration (as does 0.5 M NaCl) dissociates only the F1 histone fractions without simultaneous increase of the DNA-dependent RNA synthesis. Since similar concentrations of (NH₄)₂SO₄ increase the in vitro nuclear RNA synthesis several-fold, it can be concluded that the salt activation of RNA synthesis in isolated nuclei or chromatin is caused by the activation of RNA synthesis system and not by template derepression of chromatin. 4. 4. The pH of the salt solutions used to extract histones influences the solubility of nonhistone proteins. At pH 6.0 little nonhistone protein is extracted while at pH 8.0 considerable amount of nonhistone proteins is released into the solution.

Original language	English (US)
Pages (from-to)	202-211
Number of pages	10
Journal	BBA Section Nucleic Acids And Protein Synthesis
Volume	228
Issue number	1
DOIs	https://doi.org/10.1016/0005-2787(71)90560-0
State	Published - Jan 1 1971

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General Medicine

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title = "Proteins of chromatin in template restriction. I. RNA synthesis in vitro",

abstract = "1. 1. Selective extraction of the very lysine-rich histone fraction F1 does not increase the activity of chromatin as a template for RNA synthesis in vitro. Removal of either arginine-rich histones F2a and F3 or lysine-rich histones F2b (together with F1) results in an increase of the template activity of chromatin. 2. 2. (NH4)2SO4 removes selectively the lysine-rich histone fractions F1 and F2b. Even in 1.0 M (NH4)2SO4 the arginine-rich histones F2a and F3 remain associated with chromatin. 3. 3. (NH4)2SO4 in 0.2 M concentration (as does 0.5 M NaCl) dissociates only the F1 histone fractions without simultaneous increase of the DNA-dependent RNA synthesis. Since similar concentrations of (NH4)2SO4 increase the in vitro nuclear RNA synthesis several-fold, it can be concluded that the salt activation of RNA synthesis in isolated nuclei or chromatin is caused by the activation of RNA synthesis system and not by template derepression of chromatin. 4. 4. The pH of the salt solutions used to extract histones influences the solubility of nonhistone proteins. At pH 6.0 little nonhistone protein is extracted while at pH 8.0 considerable amount of nonhistone proteins is released into the solution.",

author = "Spelsberg, {T. C.} and Hnilica, {L. S.}",

note = "Funding Information: The authorasr ei ndebtedto Mrs. CatherineS ervanceM iss SharonT ankersley, and Mr. Rod Drake for their valuabltee chnicaal ssistancteh roughoutth eses tudies. Appreciationis extendedto Dr. John A. Wilhelm for his advicea nd criticismin the preparatioonf this manuscriptT.h e experimentawlo rk was supportedb y an American Cancer Society Grant No. E388, U.S. Public Health ServiceG rant No. CA-07746, The Robert A. Welch FoundationG rant No. G-I38, and an Institutional Grant No. FR 55II-IN-87.",

year = "1971",

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doi = "10.1016/0005-2787(71)90560-0",

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T1 - Proteins of chromatin in template restriction. I. RNA synthesis in vitro

AU - Spelsberg, T. C.

AU - Hnilica, L. S.

N1 - Funding Information: The authorasr ei ndebtedto Mrs. CatherineS ervanceM iss SharonT ankersley, and Mr. Rod Drake for their valuabltee chnicaal ssistancteh roughoutth eses tudies. Appreciationis extendedto Dr. John A. Wilhelm for his advicea nd criticismin the preparatioonf this manuscriptT.h e experimentawlo rk was supportedb y an American Cancer Society Grant No. E388, U.S. Public Health ServiceG rant No. CA-07746, The Robert A. Welch FoundationG rant No. G-I38, and an Institutional Grant No. FR 55II-IN-87.

PY - 1971/1/1

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N2 - 1. 1. Selective extraction of the very lysine-rich histone fraction F1 does not increase the activity of chromatin as a template for RNA synthesis in vitro. Removal of either arginine-rich histones F2a and F3 or lysine-rich histones F2b (together with F1) results in an increase of the template activity of chromatin. 2. 2. (NH4)2SO4 removes selectively the lysine-rich histone fractions F1 and F2b. Even in 1.0 M (NH4)2SO4 the arginine-rich histones F2a and F3 remain associated with chromatin. 3. 3. (NH4)2SO4 in 0.2 M concentration (as does 0.5 M NaCl) dissociates only the F1 histone fractions without simultaneous increase of the DNA-dependent RNA synthesis. Since similar concentrations of (NH4)2SO4 increase the in vitro nuclear RNA synthesis several-fold, it can be concluded that the salt activation of RNA synthesis in isolated nuclei or chromatin is caused by the activation of RNA synthesis system and not by template derepression of chromatin. 4. 4. The pH of the salt solutions used to extract histones influences the solubility of nonhistone proteins. At pH 6.0 little nonhistone protein is extracted while at pH 8.0 considerable amount of nonhistone proteins is released into the solution.

AB - 1. 1. Selective extraction of the very lysine-rich histone fraction F1 does not increase the activity of chromatin as a template for RNA synthesis in vitro. Removal of either arginine-rich histones F2a and F3 or lysine-rich histones F2b (together with F1) results in an increase of the template activity of chromatin. 2. 2. (NH4)2SO4 removes selectively the lysine-rich histone fractions F1 and F2b. Even in 1.0 M (NH4)2SO4 the arginine-rich histones F2a and F3 remain associated with chromatin. 3. 3. (NH4)2SO4 in 0.2 M concentration (as does 0.5 M NaCl) dissociates only the F1 histone fractions without simultaneous increase of the DNA-dependent RNA synthesis. Since similar concentrations of (NH4)2SO4 increase the in vitro nuclear RNA synthesis several-fold, it can be concluded that the salt activation of RNA synthesis in isolated nuclei or chromatin is caused by the activation of RNA synthesis system and not by template derepression of chromatin. 4. 4. The pH of the salt solutions used to extract histones influences the solubility of nonhistone proteins. At pH 6.0 little nonhistone protein is extracted while at pH 8.0 considerable amount of nonhistone proteins is released into the solution.

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Proteins of chromatin in template restriction. I. RNA synthesis in vitro

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