Activity banding of human chromosomes as shown by histone acetylation.
The expression of genes in mammalian cells depends on many factors including position in the cell cycle, stage of differentiation, age, and environmental influences. As different groups of genes are expressed, their packaging within chromatin changes and may be detected at the chromosomal level. The organization of DNA within a chromosome is determined to a large extent by the positively charged, highly conserved histones. Histone subtypes and the reversible chemical modifications of histones have been associated with gene activity. Active or potentially active genes have been associated with hyperacetylated histones and inactive genes with nonacetylated histones. Sodium butyrate increases the acetylation levels of histones in cell cultures and acts as both an inducer of gene activity and as a cell-cycle block. We describe a method to label the interphase distribution of DNA associated with various histone acetylation stages on chromosomes. Nucleosomes from untreated and butyrate-treated HeLa cells were fractionated by their acetylation level and the associated DNA labeled, and hybridized to normal human chromosomes. In the sodium butyrate-treated cells the resulting banding patterns of the high- and low-acetylated fractions were strikingly different. DNA from low-acetylated chromatin labeled several pericentric regions, whereas hybridization with DNA from highly acetylated chromatin resulted in a pattern similar to inverse G-bands on many chromosomes. The results from noninduced cells at both high and low acetylation levels were noticeably different from their induced counterparts. The capture and hybridization of DNA from interphase chromatin at different acetylation states provides a "snapshot" of the distribution of gene activity on chromosomes at the time of cell harvest.
Breneman
J W
JW
Bio-Rad Laboratories, Life Sciences Group, 2000 Alfred Nobel Drive, Hercules, CA 94547, USA.
Yau
P M
PM
Swiger
R R
RR
Teplitz
R
R
Smith
H A
HA
Tucker
J D
JD
Bradbury
E M
EM
D000107:Acetylation; D002458:Cell Fractionation; D002478:Cells, Cultured; D002501:Centrifugation, Zonal; D002843:Chromatin / Q000502:physiology; D002845:Chromatography; D002871:Chromosome Banding; D002877:Chromosomes, Human; D004247:DNA / Q000032:analysis; D017886:Durapatite; D005260:Female; D015870:Gene Expression; D006367:HeLa Cells; D006657:Histones / Q000302:isolation & purification / Q000378:metabolism*; D006801:Humans; D007621:Karyotyping; D008214:Lymphocytes / Q000166:cytology*; D008677:Metaphase; D009707:Nucleosomes / Q000502:physiology / Q000648:ultrastructure; D014960:X Chromosome
D002843:Chromatin; D006657:Histones; D009707:Nucleosomes; D004247:DNA; D017886:Durapatite