Micromechanical studies of mitotic chromosomes.
We review micromechanical experiments studying mechanoelastic properties of mitotic chromosomes. We discuss the history of this field, starting from the classic in vivo experiments of Nicklas (1983). We then focus on experiments where chromosomes were extracted from prometaphase cells and then studied by micromanipulation and microfluidic biochemical techniques. These experiments reveal that chromosomes have a well-behaved elastic response over a fivefold range of stretching, with an elastic modulus similar to that of a loosely tethered polymer network. Perturbation by microfluidic "spraying" of various ions reveals that the mitotic chromosome can be rapidly and reversibly decondensed or overcondensed, i.e., that the native state is not maximally compacted. We compare our results for chromosomes from cells to results of experiments by Houchmandzadeh and Dimitrov (1999) on chromatids reconstituted using Xenopus egg extracts. Remarkably, while the stretching elastic response of reconstituted chromosomes is similar to that observed for chromosomes from cells, reconstituted chromosomes are far more easily bent. This result suggests that reconstituted chromatids have a large-scale structure that is quite different from chromosomes in somatic cells. Finally, we discuss microspraying experiments of DNA-cutting enzymes, which reveal that the element that gives mitotic chromosomes their mechanical integrity is DNA itself. These experiments indicate that chromatin-condensing proteins are not organized into a mechanically contiguous "scaffold," but instead that the mitotic chromosome is best thought of as a cross-linked network of chromatin. Preliminary results from restriction enzyme digestion experiments indicate a spacing between chromatin "cross-links" of roughly 15 kb, a size similar to that inferred from classical chromatin loop isolation studies. These results suggest a general strategy for the use of micromanipulation methods for the study of chromosome structure.
Poirier
M G
MG
Department of Physics, University of Illinois at Chicago, Chicago, Illinois 60607, USA.
Marko
John F
JF
D000818:Animals; D001696:Biomechanical Phenomena; D002843:Chromatin / Q000187:drug effects / Q000502:physiology; D002875:Chromosomes / Q000737:chemistry / Q000187:drug effects / Q000502:physiology*; D003078:Colchicine / Q000494:pharmacology; D004262:DNA Restriction Enzymes / Q000494:pharmacology; D005057:Eukaryotic Cells / Q000166:cytology; D006801:Humans; D007477:Ions / Q000494:pharmacology; D008836:Micrococcal Nuclease / Q000494:pharmacology; D008846:Micromanipulation / Q000295:instrumentation* / Q000379:methods*; D008938:Mitosis; D008954:Models, Biological
D002843:Chromatin; D007477:Ions; D004262:DNA Restriction Enzymes; D008836:Micrococcal Nuclease; D003078:Colchicine