1022-7954/05/4104- © 2005 Pleiades Publishing, Inc.
Russian Journal of Genetics, Vol. 41, No. 4, 2005, pp. 425–429. Translated from Genetika, Vol. 41, No. 4, 2005, pp. 536–541.
Original Russian Text Copyright © 2005 by Mardanov, Krasnov, Kurshakova, Nabirochkina, Georgieva.
enchancer of yellow 2
) encodes evolutionary conservative
coactivator of RNA polymerase II. It is a ubiquitous
protein, which interacts with the transcription factor
TAF9 and is involved in regulation of transcription of a
rather large number of genes . Recently, a novel pro-
tein, E(y)2 (Sus1), which is a component of the SAGA
transcription activation complex and the nuclear pore-
associated mRNA export machinery, was described in
budding yeast .
It was demonstrated that
, as well as
some other species of
, carry the intron-con-
taining paralog of the
. The E(y)2
homologs in vertebrates and yeast are also character-
ized by intron–exon structure.
In the present study we examined the structure of the
E(y)2P protein of
, as well as the struc-
ture of its homologs
in other organisms.
is suggested to
be a retrocopy of the initial
gene, described in
the present study. It was established that during the evo-
lution E(y)2 took the functions of the initial protein as
a result of retroposition in the transcription-favorable
region. Here, we show that the initial
expressed in the testes of the
MATERIALS AND METHODS
Search for the E(y)2 homologous and analysis of
amino acid and nucleotide sequences.
orthologous sequences were found using BLAST (NCBI)
software program. Their accession numbers in the Gen-
Bank are as follows:
, AE003672; the
, AF173295; of
, AF173296; and of
, AY278445; the
, NM_144053. The
were found in the Human Genome
Sequencing Center data base [http://www.hgsc.bcm.tmc.
Alignment of the cDNA sequences was performed
using the ClustalW 1.83 software program [http://
www.genebee.msu.su/clustal]. Analysis of amino acid
sequences was carried out using the same program and
also the Multiple Sequence Alignment (InforMax) pro-
The search for the introns within the
from mouse, rat, rabbit, yeast, and human was per-
formed using the BLAT software program [http://www.
genome.ucsc.edu], while the search for the promoter
sequences was performed using the McPromoter 3.0
Synthesis of cDNA.
The cDNA samples were
obtained using 1
g of total RNA and 0.3
l of 10
oligo-dT. The volume of the reaction mixture was
adjusted to 5
l with water, the mixture was heated
C, and chilled in ice for several minutes. After the
addition of 2
l of 5
FSB (50 mM Tris–HCl, pH 8.3;
75 mM KCl; 3 mM MgCl
l 0.1 M DTT; 1
10 mM dNTPs; 0.3
l rRNAzin; and 200 units of
SuperScript RT (Gibco BRL), the mixture was incu-
bated at 42
C for 1 h.
Agarose gel was incubated for 1 h
in denaturing buffer (1.5 M NaCl; 0.5 M NaOH), and
then for 1 h in neutralizing buffer (1 M Tris–HCl;
1.5 M NaCl; 1 mM EDTA). Transfer to the Hybond N
Study of the Novel Tissue-Specific RNA Polymerase II
P. V. Mardanov
, A. N. Krasnov
, M. M. Kurshakova
E. N. Nabirochkina
, and S. G. Georgieva
1, 2, 3
Institute of Gene Biology, Russian Academy of Sciences, Moscow, 119334 Russia;
fax: (095)135-41-05; e-mail: email@example.com
Center for Medical Studies, University of Oslo, Moscow, 119334 Russia
Engelhardt Institute of Molecular Biology, Russian Academy of Sciences, Moscow, 117984 Russia; fax: (095)135-14-05
Received December 9, 2004
—It has been established that retrogenes lose introns and regulatory regions. Most of them become
pseudogenes, but some acquire tissue-speciﬁc functions. In this study, a contrary situation is described, when a
performs the main functions and is expressed in all tissues, while the
initial gene is active only in a small part of the male germ cells. It is suggested that this phenomenon resulted
from retroposition of the initial precursor gene in the transcription-suitable region of the genome.