Decreased heterogeneity of CS histone variants after hydrolysis of the ADP-ribose moiety

María Imschenetzky, Violeta Morín, Nelson Carvajal, Martín Montecino, Marcia Puchi

Resultado de la investigación: Article

11 Citas (Scopus)

Resumen

Sea urchin CS histone variants are electrophoretically heterogeneous when analyzed in two dimensional polyacrylamide gels (2D-PAGE). Previous results suggested that this heterogeneity is due to the poly (ADP-ribosylation) of these proteins. Consequently, native CS histone variants were subjected to different treatments to remove the ADP-ribose moiety. The incubation in 1 M hydroxylamine was not effective in eliminating the polymers of ADP-ribose from CS variants, and the treatment with sodium hydroxide was deleterious to the proteins. In contrast, the ADP-ribose moiety was successfully removed from the CS variants by incubation with phosphodiesterase (PDE). To eliminate contamination of CS histone variants with PDE extract, the enzyme was covalently bound to Sepharose 4B prior to its utilization. Treatment of native CS histone variants with this immobilized phosphodiesterase removed around 85% of the total ADP-ribose moiety from these proteins. After S-PDE treatment the complex electrophoretic pattern of CS histone variants in 2-D PAGE decreases to five major fractions. From these results we conclude that the electrophoretic heterogeneity of native CS histone variants is mainly due to the extent to which five main CS histone variants are poly(ADP)- ribosylated).

Idioma originalEnglish
Páginas (desde-hasta)109-117
Número de páginas9
PublicaciónJournal of Cellular Biochemistry
Volumen61
N.º1
DOI
EstadoPublished - 1996

Huella dactilar

Adenosine Diphosphate Ribose
Histones
Hydrolysis
Phosphoric Diester Hydrolases
Adenosine Diphosphate
Sodium Hydroxide
Hydroxylamine
Proteins
Sea Urchins
Electrophoresis, Gel, Two-Dimensional
Sepharose
Polymers
Contamination
Enzymes

ASJC Scopus subject areas

  • Biochemistry
  • Cell Biology
  • Molecular Biology

Citar esto

Imschenetzky, María ; Morín, Violeta ; Carvajal, Nelson ; Montecino, Martín ; Puchi, Marcia. / Decreased heterogeneity of CS histone variants after hydrolysis of the ADP-ribose moiety. En: Journal of Cellular Biochemistry. 1996 ; Vol. 61, N.º 1. pp. 109-117.
@article{b8381aaae70d47a786808d9ca3eb2100,
title = "Decreased heterogeneity of CS histone variants after hydrolysis of the ADP-ribose moiety",
abstract = "Sea urchin CS histone variants are electrophoretically heterogeneous when analyzed in two dimensional polyacrylamide gels (2D-PAGE). Previous results suggested that this heterogeneity is due to the poly (ADP-ribosylation) of these proteins. Consequently, native CS histone variants were subjected to different treatments to remove the ADP-ribose moiety. The incubation in 1 M hydroxylamine was not effective in eliminating the polymers of ADP-ribose from CS variants, and the treatment with sodium hydroxide was deleterious to the proteins. In contrast, the ADP-ribose moiety was successfully removed from the CS variants by incubation with phosphodiesterase (PDE). To eliminate contamination of CS histone variants with PDE extract, the enzyme was covalently bound to Sepharose 4B prior to its utilization. Treatment of native CS histone variants with this immobilized phosphodiesterase removed around 85{\%} of the total ADP-ribose moiety from these proteins. After S-PDE treatment the complex electrophoretic pattern of CS histone variants in 2-D PAGE decreases to five major fractions. From these results we conclude that the electrophoretic heterogeneity of native CS histone variants is mainly due to the extent to which five main CS histone variants are poly(ADP)- ribosylated).",
keywords = "chromosomal proteins, development, histone variants, poly(ADP-ribosylation), sea urchin",
author = "Mar{\'i}a Imschenetzky and Violeta Mor{\'i}n and Nelson Carvajal and Mart{\'i}n Montecino and Marcia Puchi",
year = "1996",
doi = "10.1002/(SICI)1097-4644(19960401)61:1<109::AID-JCB12>3.0.CO;2-J",
language = "English",
volume = "61",
pages = "109--117",
journal = "Journal of Cellular Biochemistry",
issn = "0730-2312",
publisher = "Wiley-Liss Inc.",
number = "1",

}

Decreased heterogeneity of CS histone variants after hydrolysis of the ADP-ribose moiety. / Imschenetzky, María; Morín, Violeta; Carvajal, Nelson; Montecino, Martín; Puchi, Marcia.

En: Journal of Cellular Biochemistry, Vol. 61, N.º 1, 1996, p. 109-117.

Resultado de la investigación: Article

TY - JOUR

T1 - Decreased heterogeneity of CS histone variants after hydrolysis of the ADP-ribose moiety

AU - Imschenetzky, María

AU - Morín, Violeta

AU - Carvajal, Nelson

AU - Montecino, Martín

AU - Puchi, Marcia

PY - 1996

Y1 - 1996

N2 - Sea urchin CS histone variants are electrophoretically heterogeneous when analyzed in two dimensional polyacrylamide gels (2D-PAGE). Previous results suggested that this heterogeneity is due to the poly (ADP-ribosylation) of these proteins. Consequently, native CS histone variants were subjected to different treatments to remove the ADP-ribose moiety. The incubation in 1 M hydroxylamine was not effective in eliminating the polymers of ADP-ribose from CS variants, and the treatment with sodium hydroxide was deleterious to the proteins. In contrast, the ADP-ribose moiety was successfully removed from the CS variants by incubation with phosphodiesterase (PDE). To eliminate contamination of CS histone variants with PDE extract, the enzyme was covalently bound to Sepharose 4B prior to its utilization. Treatment of native CS histone variants with this immobilized phosphodiesterase removed around 85% of the total ADP-ribose moiety from these proteins. After S-PDE treatment the complex electrophoretic pattern of CS histone variants in 2-D PAGE decreases to five major fractions. From these results we conclude that the electrophoretic heterogeneity of native CS histone variants is mainly due to the extent to which five main CS histone variants are poly(ADP)- ribosylated).

AB - Sea urchin CS histone variants are electrophoretically heterogeneous when analyzed in two dimensional polyacrylamide gels (2D-PAGE). Previous results suggested that this heterogeneity is due to the poly (ADP-ribosylation) of these proteins. Consequently, native CS histone variants were subjected to different treatments to remove the ADP-ribose moiety. The incubation in 1 M hydroxylamine was not effective in eliminating the polymers of ADP-ribose from CS variants, and the treatment with sodium hydroxide was deleterious to the proteins. In contrast, the ADP-ribose moiety was successfully removed from the CS variants by incubation with phosphodiesterase (PDE). To eliminate contamination of CS histone variants with PDE extract, the enzyme was covalently bound to Sepharose 4B prior to its utilization. Treatment of native CS histone variants with this immobilized phosphodiesterase removed around 85% of the total ADP-ribose moiety from these proteins. After S-PDE treatment the complex electrophoretic pattern of CS histone variants in 2-D PAGE decreases to five major fractions. From these results we conclude that the electrophoretic heterogeneity of native CS histone variants is mainly due to the extent to which five main CS histone variants are poly(ADP)- ribosylated).

KW - chromosomal proteins

KW - development

KW - histone variants

KW - poly(ADP-ribosylation)

KW - sea urchin

UR - http://www.scopus.com/inward/record.url?scp=0029989227&partnerID=8YFLogxK

U2 - 10.1002/(SICI)1097-4644(19960401)61:1<109::AID-JCB12>3.0.CO;2-J

DO - 10.1002/(SICI)1097-4644(19960401)61:1<109::AID-JCB12>3.0.CO;2-J

M3 - Article

C2 - 8726360

AN - SCOPUS:0029989227

VL - 61

SP - 109

EP - 117

JO - Journal of Cellular Biochemistry

JF - Journal of Cellular Biochemistry

SN - 0730-2312

IS - 1

ER -