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From the * Department of Biochemistry, Molecular
Biology, and Cell Biology, Northwestern University, Evanston, and
Northwestern University Medical School and the
Children's Memorial Institute for Education and Research, Chicago, Illinois.
Present address: Department of Biology, Emory
University, 1510 Clifton Rd, Atlanta, GA 30322.
Present address: Dana-Farber Cancer Institute
and Department of Genetics, Harvard Medical School, Boston, MA 02115.
| Correspondence to: Erwin Goldberg, Department of Biochemistry, Molecular Biology, and Cell Biology, Northwestern University, 2153 Sheridan Road, Evanston, IL 60208 (fax: 847-467-1380; e-mail: erv{at}northwestern.edu). |
| Received for publication April 11, 2003; accepted for publication June 12, 2003. |
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Abstract |
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Key words: Transcription, mRNA stability, protein stability
Other possible mechanisms for LDH-C4 persistence into later germ cell types are mRNA and/or protein stability. Mouse ldhc mRNA is extraordinarily stable compared with mRNA from other genes (Salehi-Ashtiani and Goldberg, 1993). No degradation of mldhc mRNA was observed after incubation of dissociated germ cells in the presence of actinomycin-D for 24 hours. For comparison, c-fos mRNA was shown to degrade rapidly, and ß-tubulin mRNA had a half-life of 10 hours.
For the present work, we demonstrated binding to the mldhc promoter by the transcription factor YY1. We also investigated the effect of 5' and 3' mldhc genomic sequences on reporter gene localization in the testes of transgenic mice. Neither the addition of approximately 4500 bp of 5' flanking DNA to the 100-bp core promoter nor the inclusion of a mldhc 3' untranslated region fragment containing the native polyadenylation signal allowed reporter expression in postmeiotic cell types. We also show that most native mldhc mRNA localizes to meiotic cells, with only a low level of postmeiotic mRNA present.
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Materials and Methods |
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TopAbstractMaterials and MethodsResultsDiscussionReferences |
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Generation of Labeled Polymerase Chain Reaction Probes
Prior to setting up polymerase chain reaction (PCR) reactions, 50 pmol of
the 5' oligonucleotide primer was end labeled by incubation with T4
polynucleotide kinase in the presence of 
-[32P]ATP. Free
nucleotides were removed by passage over a G-25 Sephadex spin column
(Pharmacia, Peapack, NJ). The labeled oligonucleotide was used in a standard
PCR reaction that contained PCR buffer (10 mM Tris-HCl [pH 8.3], 50 mM KCl,
1.5 mM MgCl2, and 0.01% [w/v] gelatin), 200 µM dNTPs, 50 pmol of
the 3' oligonucleotide primer, 2.5 U AmpliTaq DNA polymerase
(PE Biosystems, Foster City, Calif), and 20 ng plasmid DNA template containing
the mldhc promoter fragment. The oligonucleotide pairs used were
sense, MC -311 to -291 (5'-CCTACACACAGATGTAAGGGC-3') and
antisense, MC -221 to -241 (5'-TTAAAGGCCACTTTTCTGCTG-3'); sense,
MC -272 to -252 RI (5'-ccggaattcCTCCTCAAACTCAAAGTT CTG-3') and
antisense, MC -181 to -201 (5'-GACCACACAGAAGATGGCAGC-3'); and
sense, MC -239 to -219 (5'-GCAGAAAAGTGGCCTTTAAAG-3') and
antisense, MC -147 to -167 (5'-CCAAGGCCTCTAGTGTAAGAT-3').
Lowercase letters indicate nonhomologous sequence. Cycles were as follows:
94°C for 5 minutes; 25 cycles of 94°C for 1 minute, 55°C for 1
minute, and 72°C for 1 minute; and 1 cycle of 72°C for 10 minutes. The
labeled PCR product was purified by electrophoresis on a 1% agarose gel. The
desired band was excised and recovered by spinning through glass wool,
extracting with 25 : 24 : 1 phenol : chloroform : isoamyl alcohol, extracting
with 24 : 1 chloroform : isoamyl alcohol, and ethanol precipitation. The
pellet was washed briefly with 70% ethanol, air dried, and resuspended in 10
mM Tris-HCl (pH 8.0). The specific activity of probes was determined by
scintillation counting.
Generation of Oligonucleotide Probes
Complementary oligonucleotides were annealed in STE (100 mM NaCl, 10 mM
Tris-HCl [pH 8.0], and 1 mM EDTA [pH 8.0]) by heating to 95°C for 5
minutes and then cooling to room temperature over several hours.
Oligonucleotide pairs were labeled by incubation with Klenow fragment for 15
minutes at 25°C in the presence of 30 µCi of
-[32P]dCTP and 33 µM each of unlabeled dATP, dGTP, and
dTTP. Unlabeled dCTP was then added to a final concentration of 33 µM, and
the reactions were incubated for an additional 10 minutes at 25°C.
Unincorporated nucleotides were removed by passage over a G-50 Sephadex spin
column (Pharmacia). The specific activity of each labeled probe was determined
by scintillation counting.
Electrophoretic Mobility Shift Assays
Mouse testis or liver nuclear extract (7.5 µg) was incubated on ice for
15 minutes in the presence of binding buffer (13 mM HEPES [pH 7.9], 60 mM KCl,
0.13 mM EDTA, 2 mM DTT, 10% glycerol, and 0.2 mM PMSF), 1 µg poly dI-dC,
and 1 µg boiled salmon sperm DNA. The KCl present in the nuclear extract
was taken into account when calculating the final KCl concentration. A probe
(50 000 cpm) was added, and the reactions were incubated on ice for an
additional 30 minutes. Reactions were loaded onto 4% nondenaturing
polyacrylamide gels that had been prerun at 150 V at 4°C for 45 minutes.
Reactions were electrophoresed at 175 V at 4°C and then blotted onto paper
support, dried, and exposed to film at -80°C for 5 to 24 hours. Anti-YY1
immunoserum (Santa Cruz Biotechnology, Santa Cruz, Calif) was added to some
reactions. Immunoserum was either added initially or with the labeled probe.
No difference in results were seen.
Transgenic Mice Construction
The constructs used to generate transgenic animals are shown in
Figure 1. A 712-bp fragment of
the mouse ldhc promoter, including 300 bp of 5' artifactual
DNA, was amplified by PCR using the primers sense, RI/SacI 5'
(5'-ggaattcGAGCTCTGGG GGTGCTGGTTA-3') and antisense, HPAS
(5'-ATAACTGTTG GGTCCAGGAGCCAACAGTTATA-3'). Lowercase letters
indicate nonhomologous sequences incorporating an EcoRI restriction
site. The amplimer extended from a SacI site at -702 bp 5' to
the transcription start site to +10 bp 3' to the transcription start
site. The amplimer was digested with EcoRI and ligated into the
EcoRI and EcoRV sites of pBluescript II KS+ in which the
HincII, SacI, SalI, and AccI restriction
sites had been deleted. This construct was digested with SacI and
AccI, the single-stranded overhangs were blunted with mung bean
nuclease (New England Biolabs, Beverly, Mass), and the construct religated,
eliminating the 300 bp of artifactual DNA. The construct was digested with
EcoRI and XhoI, and the resulting promoter fragment was
cloned into the EcoRI and XhoI sites of pNAssß
(Clontech, Palo Alto, Calif), which resulted in pWildType (pWT)
(Figure 1A). The YY1 site at
-198 to -191 was mutated by converting nucleotides CAT to ACG using the USE
Mutagenesis Kit (Pharmacia), according to manufacturer's instructions, which
resulted in pWT mut YY1 (Figure
1B). A 722-bp fragment of the mouse ldhc 3'
untranslated region, including the native polyadenylation site, was amplified
using primers MCpoly-Asense,
5'-cgggatccACTCGCCACCTTCGACCGTGTGAC-3' and McpolyAantisense,
5'-cgggatccGTCATGTTTCACCTGCAT GTATGCC-3'. Lowercase letters
indicate nonhomologous sequence incorporating a BamHI restriction
site. The PCR product was gel-purified and digested with BamHI. pWT
was digested with BamHI, the 200-bp SV-40 polyadenylation site was
discarded, and the 722-bp mldhc 3' untranslated region PCR
amplimer was inserted in its place, which resulted in pWT 3' UTR
(Figure 1C). An approximately
4500-bp fragment of the mldhc 5' genomic sequence was excised
from a PAC clone that contained mldhc 5' flanking sequence and
exons 1 to 5 by digestion with EcoRI and KpnI. pWT 3'
UTR was digested with EcoRI and KpnI and the 5'
flanking fragment was added, which resulted in pWT 5'
(Figure 1D). The PCR-produced
portions of all constructs were sequenced on an Applied Biosystems ABI310
sequencer (ABI, Foster City, Calif) using BigDye chemistry to confirm that no
mutations had been introduced.
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Immunohistochemistry
A modification of the method described by Morales and Hecht
(1994) was used. Paraffin
embedded sections were deparaffinized by incubating 2 times for 5 minutes in
xylene and rehydrated by incubating 2 times for 3 minutes in 100% ethanol, 3
minutes in 95% ethanol, 20 minutes in 70% ethanol that contained 1%
H2O2 (to inactivate endogenous peroxidase activity), 20
minutes in 70% ethanol saturated with Li2CO3 (to
neutralize picric acid from the fixative), 3 minutes in 50% ethanol, 10
minutes in ddH2O, 10 minutes in PBS (pH 7.2), and 5 minutes in 300
mM glycine in PBS. The remainder of the immunostaining was done using reagents
from the Zymed Histostain-plus kit (Zymed Laboratories Inc, South San
Francisco, Calif). All operations were done at room temperature, except where
indicated. Tissue sections were encircled with PAP Pen (Research Products
International, Mount Prospect, Ill), to provide a well for incubations.
Sections were blocked for 10 minutes with blocking solution and then incubated
overnight at 4°C with primary antiserum diluted in PBS. Sections were
washed 2 times for 5 minutes and 1 time for 10 minutes in PBS. Sections were
incubated for 30 minutes with secondary antiserum and then washed 2 times for
5 minutes and 1 time for 10 minutes in PBS. Sections were incubated for 10
minutes with enzyme conjugate, followed by 2 times for 5 minutes and 1 time
for 10 minutes in PBS. Staining was visualized by incubation with
3,3'-diaminobenzidine (DAB)/chromogen for 1-3 min. Slides were then
rinsed well with ddH2O, counterstained for 4 minutes with
hematoxylin, rinsed briefly in 2 changes of ddH2O, dipped 5 times
into acid alcohol (1% HCl in 70% ethanol), rinsed in running tap water,
incubated for 30 seconds in Li2CO3-saturated
ddH2O, and rinsed again in ddH2O. Sections were
dehydrated by incubation for 3 minutes in 50% ethanol, 3 minutes in 70%
ethanol, 3 minutes in 95% ethanol, and 2 times for 3 minutes in 100% ethanol.
Sections were incubated for 3 minutes in xylene, air dried, and mounted with
Cytoseal (VWR Scientific).
In situ hybridization
In situ hybridization was done as described elsewhere
(Lin et al, 1997), using
mldhc cDNA as the probe.
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Results |
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YY1 Is the Factor that Recognizes the -203 to -173 Region
Computer analysis of the sequence covered by mutations 2, 3, and 4 (GCG
package, Madison, Wis) revealed a near-consensus recognition site for the
transcription factor YY1 (Hariharan et al,
1991; reviewed in Shi et al,
1997), from -198 to -191
(Figure 3B, underlined). To
investigate the possibility that YY1 was responsible for the shifted complexes
in the EMSAs, a Southwestern blot was done using the wild type -203 to -173 bp
probe and one in which the CAT within the YY1 site was mutated to ACG
(Figure 4). The wild-type probe
was bound by a factor of 65 kD, the molecular weight of YY1
(Figure 4, lane 1), whereas the
mutant probe was not (Figure 4,
lane 2).
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To confirm that YY1 was the protein binding the -203 to -173 bp probe, a
supershift analysis was done. Anti-YY1 immunoserum was added to EMSAs using
mouse testis or liver (representative somatic tissue) nuclear extract. Because
YY1 is ubiquitously expressed (Shi et al,
1997), we wanted to determine whether the binding activity
observed using testis nuclear extract was also present in somatic tissue.
Binding to the -203 to -173 probe was observed in reactions that contained
either testis or liver nuclear extract
(Figure 5A and B,
respectively). The addition of -YY1 immunoserum resulted in the
elimination of the shifted complex and the appearance of 2 supershifted bands
with lower mobility (Figure 5A and
B, arrows). The addition of a 100-fold excess of unlabeled -203 to
-173 bp probe eliminated the shifted bands (in a normal exposure; see
Figure 5 legend). However, the
addition of excess unlabeled probe containing the CAT 
ACG mutation
within the YY1 binding site mentioned above was unable to compete away the
shifted complex. The YY1 antiserum was validated by doing a Western blot on
testis nuclear extract (Figure
5C). A single band of the expected size, 65 kD, was observed,
demonstrating that the antiserum recognizes only YY1 in testis extract.
An mldhc Promoter Fragment Extending to -421 bp Regulates the
Expression of ß-Galactosidase Only in Pachytene Spermatocytes
Transgenic mouse lines were generated from 2 constructs, to investigate the
physiological relevance of the observed in vitro binding of the mldhc
promoter by YY1. The first construct contained the mldhc promoter
from -421 to +10 bp fused to the bacterial lac Z gene
(figure 1A). The second
construct was identical, except for the 3-bp CAT ACG mutation within
the YY1 site that eliminated the ability of the site to compete for binding in
the gel-shift assays (Figure
1B). Testes from male offspring of founder mice were fixed,
embedded in paraffin, sectioned, and labeled with -ß-galactosidase
immunoserum. In transgenic mice harboring the wild-type construct,
ß-galactosidase was detected only in pachytene primary spermatocytes
(Figure 6A), which is identical
to the localization reported for a promoter fragment extending to the
KpnI site at -83 bp (Li et al,
1998). Native expression also begins at the pachytene cell stage,
but LDH-C4 appears to increase during all later germ cell stages
(Hintz and Goldberg, 1977)
(Figure 6D).
ß-galactosidase localization in lines harboring the 3-bp YY1 mutation
construct was identical to that of the wild-type construct
(Figure 6B).
The Addition of 5' and 3' Flanking Sequences Does Not
Extend Reporter Expression to Postmeiotic Cell Types
The lack of reporter expression in postmeiotic cell types of mice bearing
the -421 to +10 bp promoter fragment led to 2 hypotheses. First, regulatory
elements 5' to -421 bp or in the 3' untranslated region could be
necessary for transcription in postmeiotic cell types. Second, mldhc
may not be transcribed in postmeiotic cell types, but, instead, the mRNA
persists because of mRNA stability conferred by sequence in the 3'
untranslated region. To investigate these hypotheses, an additional
approximately 4500 bp of genomic DNA was added 5' to the AccI
site at -421 bp, and a 722-bp fragment of mldhc 3' untranslated
region containing the native polyadenylation site was substituted for the
SV-40 polyadenylation site present in the expression vector
(Figure 1D). As was observed
with the previous constructs, strong ß-galactosidase expression was
observed in pachytene cells. No ß-galactosidase was detected in
postmeiotic cell types (Figure
6C) above the background observed in nontransgenic control animals
(Figure 6E). A similar reporter
expression pattern was observed for mice bearing a transgene containing the
mldhc 3' untranslated region and polyadenylation site but not the
4500-bp 5' sequence (Figure
1C; immunohistochemistry data not shown). None of the transgenic
lines exhibited a high level of postmeiotic reporter expression, as is seen
with native LDH-C4 expression
(Figure 6D). These results
suggest that, in the context of the integrated transgenes, the additional
5' and 3' sequences neither permit postmeiotic transcription nor
confer stability on the chimeric mRNA.
Mouse ldhc mRNA Is Present Predominately in Meiotic Cell Types
The apparent discrepancy between localization of native LDH-C4
and ß-galactosidase expressed from our transgenic constructs suggested
that the increased level of LDH-C4 in postmeiotic cell types may
result from mechanisms distinct from increased mldhc mRNA abundance.
In situ hybridization was done on mouse testis sections, using mldhc
cDNA as the probe, to investigate this possibility. The hybridization signal
was predominately meiotic (Figure 7A and
B, periphery of tubules), whereas a lower level of mRNA was
detectable in postmeiotic cells at higher magnification
(Figure 7B, center of tubules).
The mldhc mRNA abundance profile throughout spermatogenesis closely
matches the level of ß-galactosidase reporter seen at each stage.
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Discussion |
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YY1 has been implicated in the regulation of the testis-specific proacrosin gene (Schulten et al, 1999, 2001) which, like mldhc, is first transcribed in pachytene spermatocytes (Kashiwabara et al, 1990). The cores of the YY1 sites in both promoters are of identical sequence, GCCATNTT, although the site in the proacrosin promoter is on the minus strand and the site in the mldhc promoter is on the plus strand. The YY1 binding site in the proacrosin promoter is located within the first intron. Consensus YY1 binding sites have been found in the promoters of the testis-specific protamine 1, 2, and 3 genes and the transition protein 1 and 2 genes (Johnson et al, 1988; Kleene et al, 1990; Yelick et al, 1991; Schluter et al, 1996). However, the consensus sequence used for the search (CCATNT) was sufficiently simple that YY1 sites were found in all promoters examined. Whether YY1 plays a role in the regulation of any of these genes will have to be determined experimentally.
The substitution of the mldhc polyadenylation site and 3' untranslated region for the SV-40 polyadenylation site present in the reporter construct did not allow ß-galactosidase expression in postmeiotic germ cells. In addition, the majority of mldhc mRNA observed in the in situ hybridization experiment was in meiotic cell types, whereas a lower level was detected in postmeiotic cells. These results suggest that the mechanism for conferring stability on mldhc mRNA is not contained within the 3' untranslated region. The mldhc mRNA was found to be very stable, with no measurable degradation after a 24-hour incubation in isolated germ cells that had been treated with actinomycin-D, compared with ß-tubulin mRNA which, had a half-life of 10 hours, and c-fos mRNA, which decayed rapidly (Salehi-Ashtiani and Goldberg, 1993). It is possible that the postmeiotic mRNA results from transcription and not mRNA stability, but the fact that none of the 3 mldhc promoter fragments assayed to date regulates expression of the ß-galactosidase reporter in postmeiotic cells suggests that this is not the case. Formal proof of mldhc transcriptional activity in postmeiotic cell types is difficult to obtain without a stable cell line for such measurements. Examples of other testis-specific genes in which mRNA is stored for later translation include proacrosin (Florke et al, 1983), phosphoglycerate kinase-2 (Gold et al, 1983), protamine 1 (Kleene et al, 1984; Braun et al, 1989), and transition protein 1 (Yelick et al, 1989). For mldhc, however, the fact that mldhc mRNA decreases in abundance postmeiotically whereas LDH-C4 expression increases suggests that protein stability plays the major role in postmeiotic LDH-C4 accumulation. LDH-C4 is extremely heat-stable, retaining 65% activity after incubation at 65°C for 20 minutes (Goldberg, 1972). The LDH-C4 half-life at 65°C is 43 minutes, giving it a much greater heat stability than LDH-B4 (heart), which has a half-life of 10 minutes, and LDH-A4 (muscle), which has a half-life of 9 minutes under the same conditions. The heat stability of LDH-C4 is used to purify it from the other LDH isozymes. LDH-C4 activity persists after heat treatment, while the other LDH isozymes are inactivated.
The data presented here, along with those of previously published reports, suggest the following model for LDHC4 expression. Testis specificity is maintained by a relatively short core promoter sequence (Zhou et al, 1994; Zhou and Goldberg, 1996; Li et al, 1998), and LDH-C4 accumulation in postmeiotic germ cells is accomplished through mRNA (present article; Salehi-Ashtiani and Goldberg, 1993) and protein (Goldberg, 1972) stability. YY1 may also play a role in mldhc regulation through the control of chromatin accessibility.
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Acknowledgments |
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Footnotes |
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References |
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TopAbstractMaterials and MethodsResultsDiscussionReferences |
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Bunick D, Johnson PA, Johnson TR, Hecht NB. Transcription of the
testis-specific mouse protamine 2 gene in a homologous in vitro transcription
system. Proc Natl Acad Sci USA. 1990; 87:891-895.
Florke S, Phi-van L, Muller-Esterl W, Scheuber HP, Engel W. Acrosin in the spermiohistogenesis of mammals. Differentiation. 1983; 24:250-256.[Medline]
Gold B, Fujimoto H, Kramer JM, Erickson RP, Hecht NB. Haploid accumulation and translational control of phosphoglycerate kinase-2 messenger RNA during mouse spermatogenesis. Dev Biol. 1983; 98:392-399.[Medline]
Goldberg E. Amino acid composition and properties of crystalline
lactate dehydrogenase X from mouse testes. J Biol
Chem. 1972; 247:2044-2048.
Gorski K, Carneiro M, Schibler U. Tissue-specific in vitro transcription from the mouse albumin promoter. Cell. 1986; 47:767-776.[Medline]
Hariharan N, Kelley DE, Perry RP. Delta, a transcription factor
that binds to downstream elements in several polymerase II promoters, is a
functionally versatile zinc finger protein. Proc Natl Acad Sci
USA. 1991; 88:9799-9803.
Hintz M, Goldberg E. Immunohistochemical localization of LDH-x during spermatogenesis in mouse testes. Dev Biol. 1977; 57:375-384.[Medline]
Johnson PA, Peschon JJ, Yelick PC, Palmiter RD, Hecht NB. Sequence homologies in the mouse protamine 1 and 2 genes. Biochim Biophys Acta. 1988; 950:45-53.[Medline]
Kashiwabara S, Arai Y, Kodaira K, Baba T. Acrosin biosynthesis in meiotic and postmeiotic spermatogenic cells. Biochem Biophys Res Commun. 1990; 173:240-245.[Medline]
Kleene KC, Distel RJ, Hecht NB. Translational regulation and deadenylation of a protamine mRNA during spermiogenesis in the mouse. Dev Biol. 1984; 105:71-79.[Medline]
Kleene KC, Gerstel J, Shih D. Nucleotide sequence of the gene encoding mouse transition protein 2. Gene. 1990; 95:301-302.[Medline]
Li S, Zhou W, Doglio L, Goldberg E. Transgenic mice demonstrate a
testis-specific promoter for lactate dehydrogenase, LDHC. J Biol
Chem. 1998; 273:31191-31194.
Lin J, Poole J, Linzer DI. Two novel members of the
prolactin/growth hormone family are expressed in the mouse placenta.
Endocrinology. 1997; 138:5535-5540.
Morales CR, Hecht NB. Poly(A)+ ribonucleic acids are enriched in spermatocyte nuclei but not in chromatoid bodies in the rat testis. Biol Reprod. 1994; 50:309-319.[Abstract]
Salehi-Ashtiani K, Goldberg E. Differences in regulation of testis specifc lactate dehydrogenase in rat and mouse occur at multiple levels. Mol Reprod Dev. 1993; 35:1-7.[Medline]
Schluter G, Celik A, Obata R, Schlicker M, Hofferbert S, Schlung A, Adham IM, Engel W. Sequence analysis of the conserved protamine gene cluster shows that it contains a fourth expressed gene. Mol Reprod Dev. 1996; 43:1-6.[Medline]
Schulten HJ, Engel W, Nayernia K, Burfeind P. Yeast one-hybrid assay identifies YY1 as a binding factor for a proacrosin promoter element. Biochem Biophys Res Commun. 1999; 257:871-873.[Medline]
Schulten HJ, Nayernia K, Reim K, Engel W, Burfeind P. Assessment of promoter elements of the germ cell-specific proacrosin gene. J Cell Biochem. 2001; 83:155-162.[Medline]
Shi Y, Lee JS, Galvin KM. Everything you have ever wanted to know about Yin Yang 1. Biochim Biophys Acta. 1997; 1332:F49-F66.[Medline]
Thomas MJ, Seto E. Unlocking the mechanisms of transcription factor YY1: are chromatin modifying enzymes the key? Gene. 1999; 236:197-208.[Medline]
Yelick PC, Kozak C, Kwon YK, Seldin MF, Hecht NB. The mouse transition protein 1 gene contains a B1 repetitive element and is located on chromosome 1. Genomics. 1991; 11:687-694.[Medline]
Yelick PC, Kwon YH, Flynn JF, Borzorgzadeh A, Kleene KC, Hecht NB. Mouse transition protein 1 is translationally regulated during the post-meiotic stages of spermatogenesis. Mol Reprod Dev. 1989; 1:193-200.[Medline]
Zhou W, Goldberg E. A dual-function palindromic sequence regulates testis-specific transcription of the mouse lactate dehydrogenase c gene in vitro. Biol Reprod. 1996; 54:84-90.[Abstract]
Zhou W, Xu J, Goldberg E. A 60-bp core promoter sequence of murine lactate dehydrogenase C is sufficient to direct testis-specific transcription in vitro. Biol Reprod. 1994; 51:425-432.[Abstract]
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