L. Heliot et al., ELECTRON TOMOGRAPHY OF METAPHASE NUCLEOLAR ORGANIZER REGIONS - EVIDENCE FOR A TWISTED-LOOP ORGANIZATION, Molecular biology of the cell, 8(11), 1997, pp. 2199-2216
Metaphase nucleolar organizer regions (NORs), one of four types of chr
omosome bands, are located on human acrocentric chromosomes. They cont
ain r-chromatin, i.e., ribosomal genes complexed with proteins such as
upstream binding factor and RNA polymerase I, which are argyrophilic
NOR proteins. Immunocytochemical and cytochemical labelings of these p
roteins were used to reveal r-chromatin in situ and to investigate its
spatial organization within NORs by confocal microscopy and by electr
on tomography. For each labeling, confocal microscopy revealed small a
nd large double-spotted NORs and crescent-shaped NORs. Their internal
three-dimensional (3D) organization was studied by using electron tomo
graphy on specifically silver-stained NORs. The 3D reconstructions all
ow us to conclude that the argyrophilic NOR proteins are grouped as a
fiber of 60-80 nm in diameter that constitutes either one part of a tu
rn or two or three turns of a helix within small and large double-spot
ted NORs, respectively. Within crescent-shaped NORs, virtual slices re
veal that the fiber constitutes several longitudinally twisted loops,
grouped as two helical 250- to 300-nm coils, each centered on a nonarg
yrophilic axis of condensed chromatin. We propose a model of the 3D or
ganization of r-chromatin within elongated NORs, in which loops are tw
isted and bent to constitute one basic chromatid coil.