Disruption of topoisomerase II perturbs pairing in drosophila cell culture.
Homolog pairing refers to the alignment and physical apposition of homologous chromosomal segments. Although commonly observed during meiosis, homolog pairing also occurs in nonmeiotic cells of several organisms, including humans and Drosophila. The mechanism underlying nonmeiotic pairing, however, remains largely unknown. Here, we explore the use of established Drosophila cell lines for the analysis of pairing in somatic cells. Using fluorescent in situ hybridization (FISH), we assayed pairing at nine regions scattered throughout the genome of Kc167 cells, observing high levels of homolog pairing at all six euchromatic regions assayed and variably lower levels in regions in or near centromeric heterochromatin. We have also observed extensive pairing in six additional cell lines representing different tissues of origin, different ploidies, and two different species, demonstrating homolog pairing in cell culture to be impervious to cell type or culture history. Furthermore, by sorting Kc167 cells into G1, S, and G2 subpopulations, we show that even progression through these stages of the cell cycle does not significantly change pairing levels. Finally, our data indicate that disrupting Drosophila topoisomerase II (Top2) gene function with RNAi and chemical inhibitors perturbs homolog pairing, suggesting Top2 to be a gene important for pairing.
Williams
Benjamin R
BR
Department of Genetics, Harvard Medical School, Boston, Massachusetts 02115, USA.
Bateman
Jack R
JR
Novikov
Natasha D
ND
Wu
C-Ting
CT
D000818:Animals; D018929:Cell Culture Techniques; D002453:Cell Cycle; D023902:Chromosome Pairing; D015825:Chromosomes, Fungal / Q000235:genetics*; D004250:DNA Topoisomerases, Type II / Q000235:genetics*; D004331:Drosophila melanogaster / Q000235:genetics* / Q000254:growth & development / Q000378:metabolism; D006570:Heterochromatin; D008540:Meiosis / Q000235:genetics*; D011995:Recombination, Genetic
D006570:Heterochromatin; D004250:DNA Topoisomerases, Type II