Identification of sister chromatids by DNA template strand sequences.
It is generally assumed that sister chromatids are genetically and functionally identical and that segregation to daughter cells is a random process. However, functional differences between sister chromatids regulate daughter cell fate in yeast and sister chromatid segregation is not random in Escherichia coli. Differentiated sister chromatids, coupled with non-random segregation, have been proposed to regulate cell fate during the development of multicellular organisms. This hypothesis has not been tested because molecular features to reliably distinguish between sister chromatids are not obvious. Here we show that parental 'Watson' and 'Crick' DNA template strands can be identified in sister chromatids of murine metaphase chromosomes using CO-FISH (chromosome orientation fluorescence in situ hybridization) with unidirectional probes specific for centromeric and telomeric repeats. All chromosomes were found to have a uniform orientation with the 5' end of the short arm on the same strand as T-rich major satellite repeats. The invariable orientation of repetitive DNA was used to differentially label sister chromatids and directly study mitotic segregation patterns in different cell types. Whereas sister chromatids appeared to be randomly distributed between daughter cells in cultured lung fibroblasts and embryonic stem cells, significant non-random sister chromatid segregation was observed in a subset of colon crypt epithelial cells, including cells outside positions reported for colon stem cells. Our results establish that DNA template sequences can be used to distinguish sister chromatids and follow their mitotic segregation in vivo.
Falconer
Ester
E
Terry Fox Laboratory, B.C. Cancer Agency, Vancouver, British Columbia V5Z 1L3, Canada.
Chavez
Elizabeth A
EA
Henderson
Alexander
A
Poon
Steven S S
SS
McKinney
Steven
S
Brown
Lindsay
L
Huntsman
David G
DG
Lansdorp
Peter M
PM
D000818:Animals; D002460:Cell Line; D002842:Chromatids / Q000235:genetics* / Q000378:metabolism*; D020090:Chromosome Segregation / Q000502:physiology*; D003106:Colon / Q000166:cytology; D004276:DNA, Satellite / Q000378:metabolism; D053595:Embryonic Stem Cells / Q000166:cytology; D004847:Epithelial Cells / Q000166:cytology; D005347:Fibroblasts / Q000166:cytology; D005453:Fluorescence; D017404:In Situ Hybridization, Fluorescence / Q000379:methods*; D008163:Luminescent Measurements; D008168:Lung / Q000166:cytology; D051379:Mice; D008810:Mice, Inbred C57BL; D008938:Mitosis; D008954:Models, Biological; D009928:Organ Specificity; D013379:Substrate Specificity; D016615:Telomere / Q000378:metabolism; D013698:Templates, Genetic
D004276:DNA, Satellite