Abstract
X chromosome structural abnormalities are relatively common in Turner syndrome patients, in particular X isochromosomes. Reports over the last five decades examining asynchronous DNA replication between the normal X and isochromosome have clearly established that the structurally abnormal chromosome is the inactive X chromosome (Xi). Here the organization of chromatin at a deleted X chromosome, an Xq isochromosome, and two isodicentric chromosomes were examined. Consistent with previous differential staining methods, at interphase, the X isochromosome and isodicentric X chromosomes frequently formed bipartite Barr bodies, observed by fluorescence microscopy using numerous independent bona fide markers of Xi heterochromatin. At metaphase, with the exception of the pseudoautosomal region and the duplicated locus of the macrosatellite DXZ4 (if present on the abnormal X chromosome based on break points), euchromatin markers were absent from the Xi, whereas histone variant macroH2A formed reproducible banded mirror-image chromosomes. Unexpectedly, the isodicentric chromosome in 46,X,idic(X)(q28) cells, which carry a near full-length q-arm-to-q-arm fused chromosome, showed at interphase very rare instances of Xi chromatin bodies that were separated by large distances in the nucleus. Further examination using immunofluorescence and FISH support the possibility that these rare cells may represent ones in which one half of the isodicentric chromosome is active and the other half is inactive.
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Abbreviations
- ATCC:
-
American-type culture collection
- BSA:
-
Bovine serum albumin
- BAC:
-
Bacterial artificial chromosome
- CEP-X:
-
X alpha satellite centromere probe
- DAPI:
-
4,6-diaminidino-2-phenylindole
- FBS:
-
Fetal bovine serum
- FISH:
-
Fluorescence in situ hybridization
- H2AK119Ub1:
-
Histone H2A monoubiquitylated at lysine 119
- H3K4me2:
-
Histone H3 di-methylated at lysine 4
- H3K9me3:
-
Histone H3 tri-methylated at lysine 9
- H3K27me3:
-
Histone H3 tri-methylated at lysine 27
- HMG-Y:
-
High mobility group protein Y
- HP1:
-
Heterochromatin protein 1
- idic(X):
-
Isodicentric X chromosome
- i(X):
-
Isochromosome X
- MCB:
-
Macrochromatin body
- Mb:
-
Megabase
- PBS:
-
Phosphate-buffered saline
- PBS-T:
-
Phosphate-buffered saline supplemented with Tween-20
- SMCHD1:
-
Structural maintenance of chromosomes flexible hinge domain containing 1
- WCP-X:
-
X whole chromosome paint
- WT:
-
Wild type
- Xa:
-
Active X chromosome
- Xi:
-
Inactive X chromosome
- XIC:
-
X inactivation center
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Acknowledgments
I am grateful to Dr. Huntington Willard for continued use of the cells used in this study. I thank Dr. Stuart Schwartz for the provision of T-3352 cells. This work was supported by the National Institute of General Medical Sciences at the National Institutes of Health [grant number GM117003 to B.P.C.].
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Chadwick, B.P. Characterization of chromatin at structurally abnormal inactive X chromosomes reveals potential evidence of a rare hybrid active and inactive isodicentric X chromosome. Chromosome Res 28, 155–169 (2020). https://doi.org/10.1007/s10577-019-09621-1
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DOI: https://doi.org/10.1007/s10577-019-09621-1