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Thursday April 7 Cellular Oncology 27 (2005) 105–118 105 IOS Press Thursday April 7 PREMIER LECTURE SERIES (ICCond) with a concomitant enlargement of the IC Location: PFC G07 occurs rapidly, when living cells are exposed to a hyper- osmotic medium. Cells, which were kept up to 30 min in the ICCond state, before they were released to normal PL5 09.00-09.40 osmotic conditions, retained their viability. Exposure of cells to several, short cycles of ICCond and decondensa- Professor Thomas Cremer tion yielded highly reproducible patterns of condensed Department Biology II, Ludwig and normal chromatin textures. During ICCond individ- Maximilians University, Munich, ual CTs could still be delineated as distinct objects using Germany chromosome painting. Interestingly, neighboring CTs were not fully separated from each other, but formed a 3D higher order chromatin network. The contact sites of chromatin from different CTs have not yet been identi- fied. Splicing speckles, nuclear bodies and several pro- FROM CHROMOSOME TERRITORIES TO A teins, described as markers for a nuclear matrix, were FUNCTIONAL NUCLEAR ARCHITECTURE detected within the enlarged IC. Nascent transcripts, pulse-labeled immediately prior to ICCond with BrUTP, Thomas Cremer, Heiner Albiez, Andreas Bolzer, Daniela were detected at the surface of condensed chromatin Köhler, Katrin Küpper, Stefan Müller, Michaela Neusser, streaks lining the enlarged IC. This and other experi- Lothar Schermelleh, Heinrich Leonhardt, Babett Wagler, ments argue that for transcription genes must be posi- Roman Zinner, Marion Cremer, Irina Solovei tioned in perichromatin regions lining the IC or on chro- Department Biology II, Ludwig Maximilians University, matin loops expanding into the IC. Transcription as well Munich, Grosshadernerstr. 2, 82152 Martinsried, Germany as DNA replication was inhibited during ICCond, but continued as soon as the normal chromatin condensation The chromosome territory - interchromatin compartment state was restored. We propose that CTs and the IC form (CT-IC) model argues for a functional organization of the two separate, yet structurally and functionally closely cell nucleus. Chromosome territory (CT) arrangements associated nuclear networks. Border zones of chromatin show profound differences between certain cell types. In domains, also called perichromatin regions, line the IC the spherically shaped lymphocyte nuclei gene dense and provide essential sites for transcription, co- CTs are preferentially located in the nuclear interior, transcriptional splicing, DNA replication and DNA re- while gene poor CTs are preferentially located towards pair. the nuclear periphery. In fibroblast nuclei with their flat- ellipsoidal shape, however, CTs of small chromosomes - Biography independent of their gene density - are located towards Thomas Cremer was born in 1945. From 1964 to 1970 the nuclear center and CTs of large chromosomes to- he studied human medicine at the University of Freiburg th wards the nuclear rim. Studies of other primates indicate (Germany). In the early 70 he built a laser-uv- an evolutionary conservation of gene density correlated microbeam apparatus together with his brother Christoph and size correlated, radial arrangements in the respective Cremer, presently full Professor of Applied Optics at the cell types. Using defined BACs in combination with University of Heidelberg. Their early microbeam studies multicolor 3D FISH for gene dense and gene poor chro- with living cells yielded compelling evidence for chro- mosomal sub-regions, we have started to define the 3D mosome territories in nuclei of mammalian cells. Since architecture of individual CTs and their topological inter- 1974 T.C. has led his own research group first at the actions with each other and the IC. In addition, we have University of Freiburg i. Br. and since 1978 at the Uni- established a procedure to manipulate chromatin conden- versity of Heidelberg. 1978 the brothers published the sation and the width of the IC in nuclei of living cells in a first construction proposal for a laser confocal scanning fully reversible way. Induced chromatin condensation fluorescence microscope to enable 3D-imaging of cells. 1570-5870/05/$17.00 © 2005 – IOS Press and the authors. All rights reserved 106 Thursday April 7 In 1978 Professor Cremer was a guest researcher in the However, biophysical evidence for different chromatin laboratory of Michel W. Berns, University of California, fibre structures which might equate with these concepts Irvine. From 1986 - 1988 he worked as a visiting Profes- has been lacking. Cancer is a disease of aberrant gene sor at Yale, Medical School, New Haven, together with expression. Since gene expression is regulated by chro- Laura Manuelidis and David C. Ward. During this time matin structure it is important to understand how the he contributed essentially to the development of FISH- chromatin structure can affect gene expression. protocols for the visualization of individual chromo- In normal lymphoblastoid cells we have undertaken the somes in human cells. Thereafter he worked as a lecturer first analysis of chromatin fibre structure across the hu- of human genetics at the University of Heidelberg. In man genome. By sucrose sedimentation we separated 1996 he became Professor and Chair of Anthropology compact and open chromatin fibre structures. Their dis- and Human Genetics in the Faculty of Biology, Univer- tribution was analysed by hybridisation to metaphase sity of Munich. Major topics of research: development of chromosomes (low resolution) and genomic microarrays FISH tools for interphase cytogenetics and studies of (high resolution). A whole genome microarray assem- higher order chromatin arrangements in the cell nucleus, bled from clones, spaced at ~1Mb intervals, from the as well as chromosome and nuclear architecture evolu- “golden path” used in the sequencing of the human ge- tion; comparative genomic hybridization (CGH) and nome and a chromosome 22 tiling-path array with an matrix (or array) CGH; experimental studies of the func- average resolution of 78kb were used. By utilising ge- tional compartmentalization of the cell nucleus led to the nomic microarrays of known sequence we can now relate formulation of the chromosome territory – interchro- known genomic features and gene expression levels to matin compartment model (with C. Cremer and P. Lich- the underlying chromatin structure. ter). Since 1998 the Cremer laboratory has made major We have shown that compact chromatin fibres originate efforts to develop approaches for studies of higher order from some sites of heterochromatin (C-bands), and chromatin arrangements and their dynamics in nuclei of G-bands (euchromatin), whilst open chromatin fibres living cells. correlate with regions of highest gene density, but not with gene expression. Inactive genes can be in domains of open chromatin, and active genes in regions of low PL6 09.40-10.20 gene density can be in compact chromatin. We have also shown that chromatin fibre structure impacts on further Dr Nick Gilbert levels of chromatin condensation as regions of open MRC Human Genetics Unit, chromatin fibres are cytologically decondensed and have Edinburgh, UK a distinctive nuclear organisation. We would suggest that domains of open chromatin create an environment that facilitates transcriptional activation whilst an aberrant alteration in chromatin structure will cause widespread CHROMATIN ARCHITECTURE OF THE HU- misregulation of genes. MAN GENOME Biography Human cells package their DNA into chromatin, a nu- Having studied biochemistry at The University of Edin- cleoprotein complex. In chromatin the DNA is wrapped burgh, Dr Gilbert spent a short period in Cambridge 1.75 times around a nucleosome, consisting of an oc- working on “cancer gene therapy. He completed his PhD tamer of histone proteins. Under physiological salt condi- in Edinburgh studying the higher-order structure of tions nucleosome arrays adopt the conformation of a mammalian heterochromatin. He continues to working 30nm fibre. Cellular processes such as gene transcription, chromatin research at the MRC Human Genetics Unit DNA replication and repair occur in this environment with a specific interest in mammalian higher-order chro- and the modulation of the chromatin structure is central matin fibre and how its conformation is modulated. to their regulation. The structure of the chromatin fibre is best understood at the level of the nucleosome and its References. modifications such as acetylation and methylation. How- [1] Gilbert, N. and Allan, J. (2001). Distinctive higher- ever, beyond the nucleosome itself there are other states order chromatin structure at mammalian centro- of chromatin that will influence how the underlying meres. Proc. Natl Acad. Sci. USA 98, 11949-11954. DNA sequence is read. Transcriptionally active and silent [2] Gilbert, N., Boyle, S., Sutherland, H., de Las Heras, regions of the genome are often considered to have J., Allan, J., Jenuwein, T., Bickmore, W. A. (2003) "open" and "closed" chromatin structures respectively. Thursday April 7 107 Formation of facultative heterochromatin in the ab- of the mitotic division machinery and are testing their sence of HP1. EMBO J. 22, 5540-5550. potential for formation of aneuploidy and tumor forma- [3] Gilbert, N., Gilchrist, S., Bickmore, W. A. (2004) tion. These investigations of the fundamental cell bio- Chromatin organisation in the mammalian nucleus. logical principles of nuclear architecture and genome Int. Rev. Cytol. (in press).
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