Subcellular location of enzymes involved in core histone acetylation.
Multiple enzyme forms of histone deacetylase and histone acetyltransferase exist in germinating maize embryos. We analyzed the association of the different enzymes to chromatin by ion exchange chromatography of subcellular fractions from different time points of embryo germination. The vast majority of histone deacetylase HD-1A was not bound to chromatin, since it was solubilized during chromatin isolation, regardless of its phosphorylation state and the phase of embryo germination. In contrast, HD-2 was chromatin bound during the entire germination pathway. Histone deacetylase HD-1B was present in a chromatin-bound and a soluble form; the ratio between these two forms changed during germination. Both nuclear histone acetyltransferases, HAT-A1 and HAT-A2, were tightly chromatin-bound and could only be released from chromatin by salt extraction. To test whether histone acetyltransferases or deacetylases are associated with the nuclear matrix, we analyzed nuclear matrix preparations from yeast, Physarum, and maize step by step for both enzyme activities. This analysis confirmed that part of the activity is chromatin bound, but no significant enzyme activity could be found in the final nuclear matrix, regardless of the preparation protocol. This result was further substantiated by detailed analysis of histone deacetylases and acetyltransferases during cellular fractionation and nuclear matrix preparation of chicken erythrocytes. Altogether our results suggest that the participation of these enzymes in different nuclear processes may partly be regulated by a distinct location to intranuclear components.
Grabher
A
A
Department of Microbiology, University of Innsbruck, Medical School, Austria.
Brosch
G
G
Sendra
R
R
Lechner
T
T
Eberharter
A
A
Georgieva
E I
EI
López-Rodas
G
G
Franco
L
L
Dietrich
H
H
Loidl
P
P
D000107:Acetylation; D000123:Acetyltransferases / Q000032:analysis*; D000818:Animals; D002467:Cell Nucleus / Q000201:enzymology*; D002645:Chickens; D002852:Chromatography, Ion Exchange; D002853:Chromatography, Liquid / Q000379:methods; D004912:Erythrocytes / Q000201:enzymology; D051548:Histone Acetyltransferases; D006655:Histone Deacetylases / Q000032:analysis*; D006657:Histones / Q000378:metabolism*; D008297:Male; D015530:Nuclear Matrix / Q000201:enzymology; D016839:Physarum polycephalum / Q000201:enzymology; D012441:Saccharomyces cerevisiae / Q000201:enzymology; D029701:Saccharomyces cerevisiae Proteins; D012639:Seeds / Q000201:enzymology / Q000254:growth & development; D013347:Subcellular Fractions / Q000201:enzymology*; D003313:Zea mays / Q000201:enzymology
D006657:Histones; D029701:Saccharomyces cerevisiae Proteins; D000123:Acetyltransferases; D051548:Histone Acetyltransferases; D006655:Histone Deacetylases