Abstract
The metaphase chromosome spread technique and subsequent analysis of sister chromatid cohesion is used for (clinical) diagnosis of genetic abnormalities that can cause aberrant sister chromatid cohesion. In addition, the technique can be used to assess the contribution of novel genes to the cohesion establishment and maintenance pathways. Cells are swelled in a hypotonic solution and fixed in Carnoy’s solution. Samples are then dropped onto glass slides, and the spread chromosomes are stained and visualized by microscopy. Defects in sister chromatid cohesion can be easily assessed using this method, examples of which are given.
This is a preview of subscription content, log in via an institution to check access.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Ricke RM, van Ree JH, van Deursen JM (2008) Whole chromosome instability and cancer: a complex relationship. Trends Genet 24:457–466
Weaver BA, Cleveland DW (2006) Does aneuploidy cause cancer? Curr Opin Cell Biol 18:658–667
Feeney KM, Wasson CW, Parish JL (2010) Cohesin: a regulator of genome integrity and gene expression. Biochem J 428:147–161
Michaelis C, Ciosk R, Nasmyth K (1997) Cohesins: chromosomal proteins that prevent premature separation of sister chromatids. Cell 91:35–45
Haering CH, Lowe J, Hochwagen A et al (2002) Molecular architecture of SMC proteins and the yeast cohesin complex. Mol Cell 9:773–788
Lengronne A, Katou Y, Mori S, Yokobayashi S et al (2004) Cohesin relocation from sites of chromosomal loading to places of convergent transcription. Nature 430:573–578
Darwiche N, Freeman LA, Strunnikov A (1999) Characterization of the components of the putative mammalian sister chromatid cohesion complex. Gene 233:39–47
Ciosk R, Shirayama M, Shevchenko A et al (2000) Cohesin’s binding to chromosomes depends on a separate complex consisting of Scc2 and Scc4 proteins. Mol Cell 5:243–254
Kurze A, Michie KA, Dixon SE et al (2011) A positively charged channel within the Smc1/Smc3 hinge required for sister chromatid cohesion. EMBO J 30:364–378
Whelan G, Kreidl E, Peters JM et al (2012) The non-redundant function of cohesin acetyltransferase Esco2: some answers and new questions. Nucleus 3:330–334
Zhang J, Shi X, Li Y et al (2008) Acetylation of Smc3 by Eco1 is required for S phase sister chromatid cohesion in both human and yeast. Mol Cell 31:143–151
Nishiyama T, Ladurner R, Schmitz J et al (2010) Sororin mediates sister chromatid cohesion by antagonizing Wapl. Cell 143:737–749
Losada A, Hirano M, Hirano T (1998) Identification of Xenopus SMC protein complexes required for sister chromatid cohesion. Genes Dev 12:1986–1997
Nishiyama T, Sykora MM, Huis in’t Veld PJ et al (2013) Aurora B and Cdk1 mediate Wapl activation and release of acetylated cohesin from chromosomes by phosphorylating Sororin. PNAS 110:13404–13409
Sumara I, Vorlaufer E, Stukenberg PT et al (2002) The dissociation of cohesin from chromosomes in prophase is regulated by Polo-like kinase. Mol Cell 9:515–525
Waizenegger IC, Hauf S, Meinke A et al (2000) Two distinct pathways remove mammalian cohesin from chromosome arms in prophase and from centromeres in anaphase. Cell 103:399–410
Liu H, Rankin S, Yu H (2013) Phosphorylation-enabled binding of SGO1-PP2A to cohesin protects sororin and centromeric cohesion during mitosis. Nat Cell Biol 15:40–49
Shintomi K, Hirano T (2010) Sister chromatid resolution: a cohesin releasing network and beyond. Chromosoma 119:459–467
van der Lelij P, Chrzanowska KH, Godthelp BC et al (2010) Warsaw breakage syndrome, a cohesinopathy associated with mutations in the XPD helicase family member DDX11/ChlR1. Am J Hum Genet 86:262–266
Inoue A, Li T, Roby SK, Valentine MB et al (2007) Loss of ChlR1 helicase in mouse causes lethality due to the accumulation of aneuploid cells generated by cohesion defects and placental malformation. Cell Cycle 6:1646–1654
Parish JL, Rosa J, Wang X et al (2006) The DNA helicase ChlR1 is required for sister chromatid cohesion in mammalian cells. J Cell Sci 119:4857–4865
Acknowledgements
JP is supported by a Royal Society University Research Fellowship (UF110010).
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2014 Springer Science+Business Media New York
About this protocol
Cite this protocol
Feeney, K.M., McFarlane-Majeed, L., Parish, J.L. (2014). Analyzing Sister Chromatid Cohesion in Mammalian Cells. In: Noguchi, E., Gadaleta, M. (eds) Cell Cycle Control. Methods in Molecular Biology, vol 1170. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-0888-2_32
Download citation
DOI: https://doi.org/10.1007/978-1-4939-0888-2_32
Published:
Publisher Name: Humana Press, New York, NY
Print ISBN: 978-1-4939-0887-5
Online ISBN: 978-1-4939-0888-2
eBook Packages: Springer Protocols