Lampbrush chromosomes enable study of cohesin dynamics |
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Authors: | Christopher Austin Natalya Novikova Vincent Guacci Michel Bellini |
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Institution: | (1) Cell & Developmental Biology, University of Illinois at Urbana-Champaign, Urbana, IL, USA;(2) Embryology, Carnegie Institution of Washington, Baltimore, MD, USA;(3) Department of Cell and Developmental Biology, School of Molecular and Cellular Biology, 601 South Goodwin Avenue, Room B107 CLSL, Urbana, IL 61801, USA |
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Abstract: | The lampbrush chromosomes present in the nuclei of amphibian oocytes offer unique biological approaches for study of the mechanisms
that regulate chromatin structure with high spatial resolution. We discuss fundamental aspects of the remarkable organization
and plasticity exhibited by lampbrush chromosomes. We then utilize lampbrush chromosomes to characterize the chromosomal distribution
and dynamics of cohesin, the four-protein complex (RAD21/MCD1/SCC1, SMC1, SMC3, SCC3/SA2) responsible for sister chromatid
cohesion. We find that endogenous SMC3 and newly expressed hRAD21 co-localize on chromosomal axes, sites where sister chromatids
are tightly paired. We present evidence suggesting that hRAD21 recruitment to lampbrush chromosomes is modulated by chromosomal
SMC1 and SMC3. Notably, using a technique for de novo chromosome assembly, we demonstrate that both SMC3 and hRAD21 are recruited to single, unreplicated lampbrush chromatids.
Finally, we used our novel method of analyzing the oocyte nucleus under oil combined with fluorescence recovery after photobleaching,
to provide direct evidence that cohesin is highly dynamic at discrete, condensed chromosomal regions. Collectively, these
data demonstrate that lampbrush chromosomes provide a unique and powerful tool for combining biochemical and cytological analyses
for dissection of complex chromosomal processes. |
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Keywords: | lampbrush chromosome chromatin cohesin SMC oocyte nucleus |
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