Visualizing the Spatial Relationships between Defined DNA Sequences and the Axial Region of Extracted Metaphase Chromosomes

Using fluorescence in situ hybridization to extracted metaphase chromosomes, we present visual evidence that specific human DNA sequences occupy distinctive positions with respect to the axial region of chromosomes and that the DNA is organized into loops emanating from this region. In a stretch of unique DNA on chromosome 11, large loops of DNA can be traced and one specific region associated with the axial region of the chromosome. Within rDNA, nontranscribed spacer sequences are more closely apposed to the chromosome axis than are rRNA genes. Heterochromatic and euchromatic DNAs appear to be organized into loops of similar size. We could not detect loops at centromeres; most alphoid DNA appears to remain close to the axial region.

[1]  P. Dijkwel,et al.  Permanent attachment of replication origins to the nuclear matrix in BHK-cells. , 1986, Nucleic acids research.

[2]  W. Earnshaw Mitotic chromosome structure , 1988, BioEssays : news and reviews in molecular, cellular and developmental biology.

[3]  Professor Dr. Harold Garnet Callan Lampbrush Chromosomes , 1986, Molecular Biology, Biochemistry and Biophysics.

[4]  R. D. Little,et al.  Initiation and termination of DNA replication in human rRNA genes , 1993, Molecular and cellular biology.

[5]  Y. Fukushima,et al.  Aniridia-associated cytogenetic rearrangements suggest that a position effect may cause the mutant phenotype. , 1995, Human molecular genetics.

[6]  J. Sedat,et al.  Materials and methods , 2004, Documenta Ophthalmologica.

[7]  U. K. Laemmli,et al.  Organization of the higher-order chromatin loop: specific DNA attachment sites on nuclear scaffold , 1984, Cell.

[8]  E. Bruford,et al.  A high-resolution integrated physical, cytogenetic, and genetic map of human chromosome 11: distal p13 to proximal p15.1. , 1995, Genomics.

[9]  U. K. Laemmli,et al.  Metaphase chromosome structure. Involvement of topoisomerase II. , 1986, Journal of molecular biology.

[10]  W. Earnshaw,et al.  Localization of topoisomerase II in mitotic chromosomes , 1985, The Journal of cell biology.

[11]  R. Hancock,et al.  Structural-functional organization of chromosomal DNA domains. , 1993, Cold Spring Harbor symposia on quantitative biology.

[12]  W. Earnshaw,et al.  ScII: an abundant chromosome scaffold protein is a member of a family of putative ATPases with an unusual predicted tertiary structure , 1994, The Journal of cell biology.

[13]  H. G. Callan,et al.  The Croonian Lecture, 1981 - Lampbrush chromosomes , 1982, Proceedings of the Royal Society of London. Series B. Biological Sciences.

[14]  P. Cook The nucleoskeleton and the topology of replication , 1991, Cell.

[15]  B. Trask,et al.  Mapping of human chromosome Xq28 by two-color fluorescence in situ hybridization of DNA sequences to interphase cell nuclei. , 1991, American journal of human genetics.

[16]  W. Earnshaw,et al.  The kinetochore is part of the metaphase chromosome scaffold , 1984, The Journal of cell biology.

[17]  S. Gasser,et al.  Studies on scaffold attachment sites and their relation to genome function. , 1989, International review of cytology.

[18]  J. Huberman,et al.  University of Groningen Organization of Replication of Ribosomal DNA in Saccharomyces cerevisiae , 2017 .

[19]  E. Viégas-Péquignot,et al.  Genes occupy a fixed and symmetrical position on sister chromatids , 1991, Cell.

[20]  C. D. Lewis,et al.  Higher order metaphase chromosome structure: Evidence for metalloprotein interactions , 1982, Cell.

[21]  R. Young,et al.  RNA polymerase II. , 1991, Annual review of biochemistry.

[22]  J. R. Paulson,et al.  The structure of histone-depleted metaphase chromosomes , 1977, Cell.

[23]  D. Jackson,et al.  The size of chromatin loops in HeLa cells. , 1990, The EMBO journal.

[24]  M. Buongiorno-Nardelli,et al.  A relationship between replicon size and supercoiled loop domains in the eukaryotic genome , 1982, Nature.

[25]  J. R. Paulson,et al.  Metaphase chromosome structure , 1977 .

[26]  W. Earnshaw,et al.  Mapping DNA within the mammalian kinetochore , 1993, The Journal of cell biology.

[27]  P. Jeppesen,et al.  Effects of sulphydryl reagents on the structure of dehistonized metaphase chromosomes. , 1985, Journal of cell science.

[28]  W. Earnshaw,et al.  CENP-B: a major human centromere protein located beneath the kinetochore , 1990, The Journal of cell biology.

[29]  S. Gasser,et al.  Chromosomal ARS and CEN elements bind specifically to the yeast nuclear scaffold , 1988, Cell.

[30]  H. Cooke Repeated sequence specific to human males , 1976, Nature.

[31]  U. K. Laemmli,et al.  Architecture of metaphase chromosomes and chromosome scaffolds , 1983, The Journal of cell biology.

[32]  C. Brun,et al.  Mapping of replication initiation sites in human ribosomal DNA by nascent-strand abundance analysis , 1995, Molecular and cellular biology.

[33]  U. K. Laemmli,et al.  Scaffold attachment of DNA loops in metaphase chromosomes. , 1988, Journal of molecular biology.

[34]  W. Bickmore,et al.  Factors affecting the timing and imprinting of replication on a mammalian chromosome. , 1995, Journal of cell science.

[35]  R. Zinkowski,et al.  CENP‐E, a novel human centromere‐associated protein required for progression from metaphase to anaphase. , 1991, The EMBO journal.

[36]  J. Lawrence,et al.  Dynamic changes in the higher-level chromatin organization of specific sequences revealed by in situ hybridization to nuclear halos , 1994, The Journal of cell biology.

[37]  U. K. Laemmli,et al.  Metaphase chromosome structure: Evidence for a radial loop model , 1979, Cell.

[38]  U. K. Laemmli,et al.  Metaphase chromosome structure: Bands arise from a differential folding path of the highly AT-rich scaffold , 1994, Cell.

[39]  P. Cook RNA polymerase: structural determinant of the chromatin loop and the chromosome. , 1994, BioEssays : news and reviews in molecular, cellular and developmental biology.