Bovine satellite DNA induces heterochromatinization of host chromosomal DNA in cells of trassatellite mouse embryonal carcinoma.
Embryonal teratocarcinoma F9 cells were transfected with a fragment (3.8 kb) of bovine satellite DNA IV (Sat), which is not homologous to mouse satellite DNA. FISH analysis revealed various chromosomal integration sites of integrated Sat in different transsatellite clones. After several passages, transsatellite had a tendency to spread along chromosome bearing Sat in one of the studied lines. The integrated transsatellites were enriched with prolonged single-strand DNA regions (SSR) revealed by FISH without previous chromosomal denaturation, and were unmethylated. The observed SSR are presumably supposed to represent intermediates of transsatellite DNA instability via unequal sister chromatid exchanges. DAPI staining demonstrated that the integrated Sat induced the formation of prominent ectopic neoheterochromatin blocks in regions adjacent to integrated Sat. These blocks were located exclusively between integrated Sat and centromeric heterochromatin. Thus, mouse repetitive centromeric DNA (AT-rich, DAPI-positive) "spreads" along the chromosome in response to integration of the bovine satellite GC-rich DNA. The results obtained are discussed in the context of possible position effect variegation mechanisms operating in undifferentiated cells.
Suchkova
I O
IO
Institute of Experimental Medicine RAMS, St. Petersburg.
Baranova
T V
TV
Kustova
M E
ME
Kisljakova
T V
TV
Vassiliev
V B
VB
Slominskaja
N O
NO
Alenina
N V
NV
Patkin
E L
EL
D000818:Animals; D018236:Carcinoma, Embryonal; D002417:Cattle; D045744:Cell Line, Tumor; D002875:Chromosomes / Q000235:genetics / Q000378:metabolism*; D002999:Clone Cells; D004276:DNA, Satellite; D006570:Heterochromatin / Q000235:genetics / Q000378:metabolism*; D017404:In Situ Hybridization, Fluorescence; D051379:Mice; D014162:Transfection
D004276:DNA, Satellite; D006570:Heterochromatin