ISSN 10227954, Russian Journal of Genetics, 2010, Vol. 46, No. 9, pp. 1042–1044. © Pleiades Publishing, Inc., 2010.
Original Russian Text © A.N. Shabarina, N.G. Shostak, M.V. Glazkov, 2010, published in Genetika, 2010, Vol. 46, No. 9, pp. 1175–1177.
Various regions of chromosomes are attached to the
nuclear envelope during the interphase, which ensures
a highly ordered organization of chromosomes in the
nucleus. This, in turn, is important for the “proper”
functioning of the genome, including its functions
during ontogeny. Recent data indicate that the spatial
arrangement of chromosomes in the nucleus is an ele
ment of gene expression control [1, 2]. The associa
tion (interaction) of interphase chromosomes and the
nuclear envelope via specific regions of chromosomal
DNA termed nuclear envelope DNAs (neDNAs)
plays an important role in this process.
We have a collection of chromosomal DNA frag
ments isolated from the nuclear envelopes of mouse
hepatocytes . Nuclear envelope DNAs constitute a
separate class of “structural” regions of chromosomal
DNA different from MARs/SARs (DNA fragments
isolated from the cores of the rosettelike structures
and DNA of the synaptonemal complex) . We
selected one element from this collection,
whose characteristic feature is a polypurine track
highly homologous (up to 100%) to the genomes of
various objects from microorganisms to
to humans. This evolutionarily con
served sequence is usually located in intergenic regions
of the genomes of various organisms; more rarely, it is
found in introns of genes . Our data  show that
introduction of neDNA into the construction of a
transgene (at its flanks) increases the viability of trans
genic mouse embryos by a factor of three, thereby
increasing the transgenosis efficiency. This may result
from compartmentalization of the transgene both in
chromosomes and in the interphase nucleus as a
whole, not interfering with the normal implementa
tion of the genetic program of development.
In this study, we analyzed the functional character
istics of neDNAs that are involved in preventing trans
gene heterochromatization and attempted at deter
mining their regulatory properties.
For this purpose, we used the transgenosis method
and a model system containing the
. The experimental con
struction had the following composition: 5'
, where 5'
are the ends of a
are the enhancers of the
is the enhancer of the
promoter of the
the studied sequence (
). A similar construction
not containing neDNA fragment served as a control.
To create these constructions, we used the vectors
to which the regula
tory elements of the
gene and the enhancer of
gene were cloned (the vectors were kindly
provided by P.G. Georgiev).
We used the
(carrying the gene of
nase, which allowed us to perform incision via the site
specific recombination mechanism in transgenic
). All strains were maintained at a tempera
on the standard medium. For inducing
the expression of
recombinase, second to third
instar larvae obtained by crossing the analyzed strain
strain were subjected to heat shock at
for 1.5 h on two consecutive days.
embryos were transformed using the
microinjection method according to the standard pro
tocol . The DNA of the experimental construction
dissolved in water was mixed with the DNA of a helper
) at a ratio of 5 : 1. This mixture was
embryos by means of a
micromanipulator and a microinjector (Eppendorf,
The Role of Chromosomal Regions Anchored to the Nuclear
Envelope in the Functional Organization of Chromosomes
A. N. Shabarina
, N. G. Shostak
, and M. V. Glazkov
Koltsov Institute of Developmental Biology, Russian Academy of Sciences, Moscow, 119334 Russia;
Engelhardt Institute of Molecular Biology, Russian Academy of Sciences, Moscow, 119991 Russia
Received January 28, 2010
The functional characteristics of the DNA fragments responsible for chromosome attachment to the nuclear
envelope during the interphase (neDNAs) have been studied. The neDNAs flanking the transgene have been
found to promote a steadily high rate of its expression, irrespective of the site of its insertion into the host
chromosomes. At the same time, neDNAs themselves have no transcription regulatory functions.