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Molecular Biology and Genetics |
Departments of Molecular Biotechnology [B. L., J. T. W., P. S. N.], Medicine [P. S. N.], Urology [S. W., R. V.], Microbiology [R. B.], and Pathology [L. D. T.], University of Washington, Seattle, Washington 98195; the Institute for Systems Biology, Seattle, Washington 98105 [L. H.]; and the Division of Human Biology, Fred Hutchinson Cancer Research Center, Seattle, Washington 98109 [C. F., P. S. N.]
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ABSTRACT |
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INTRODUCTION |
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In addition to a role in driving cellular proliferation, an intact androgen signaling system may also be associated with tumor suppression (4) . This dual role of androgens would not be unexpected, because androgens are responsible for differentiation of the prostate epithelium and for the regulation of specific epithelial cell functions such as the expression of PSA (5 , 6) . Several androgen-regulated genes have been demonstrated to be associated with a proliferative shut-off function in LNCaP cells and for the regulation of the cell cycle (7 , 8) . At the time of invasion or metastasis, mutations in the AR may occur (9) , suggesting that a normal AR is protective from progression. Finally, in vitro studies indicate that there may be a survival advantage in maintaining an androgen-responsive cohort of prostate tumor cells (10) . This concept has been extended to clinical medicine in which several trials suggest a benefit for an approach using intermittent rather than continuous androgen suppression in patient cohorts with hormone-responsive disease (11 , 12) .
The pivotal role of androgens in the biology and treatment of prostate cancer has led to intensive investigations designed to identify the molecular mediators of androgen action (13 , 14) . Among the genes shown to be regulated by androgens in prostate cells are several that encode enzymes belonging to the two major lipogenic pathways: fatty acid synthesis and cholesterol synthesis (15 , 16) . The regulation of cholesterol metabolism by androgens is especially intriguing because cholesterol is an essential precursor for the biosynthesis of androgens (17) . Other molecules involved in androgen metabolism and androgen action are themselves androgen regulated. For example, 17-ß-HSD, an enzyme that converts androstenedione to testosterone, is androgen regulated (18) , as is the expression of the AR itself (7 , 19) . It appears that multiple autoregulatory levels of androgen action may be operative in androgen-responsive tissues.
Our objective in this study was to identify genes that exhibit transcriptional regulation by androgens in human prostate cells. We hypothesized that such genes could be direct mediators of androgen action and that the characterization of these genes and their cognate proteins would provide insights into the mechanisms of androgen-dependent and androgen-independent cellular growth. We used cDNA microarrays comprised of cDNAs derived from human prostate tissues to quantitate transcripts expressed in the androgen-sensitive LNCaP prostate tumor cell line under conditions of androgen starvation or androgen stimulation. Here we report the cDNA cloning, chromosomal mapping, genomic structure, and expression profile of a novel gene, PSDR1, that exhibits homology to the family of SDR enzymes. PSDR1 is the first SDR shown to be predominantly expressed in normal and neoplastic prostate tissue. We hypothesize that PSDR1 may play a role in steroid hormone metabolism in prostate cells and thus may be an ideal target for modulating hormone-mediated prostate cancer growth.
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MATERIALS AND METHODS |
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Probe Construction and Microarray Hybridization.
Total RNA was isolated from LNCaP cells after 72 hrs of androgen
depletion or supplementation using TRIzol (Life Technologies) according
to the manufacturer’s directions. Fluorescence-labeled probes were
made from 30 µg total RNA in a reaction volume of 20 µl containing
1 µl of anchored oligo-dT primer (Amersham), 0.05 mM
Cy3-dCTP or Cy5-dCTP (Amersham); 0.05 mM dCTP; 0.1
mM each dGTP, dATP, dTTP; and 200 units of Superscript II
reverse transcriptase (Life Technologies). Reactants were incubated at
42°C for 120 min followed by heating to 94°C for 3 min. Unlabeled
RNA was hydrolyzed by the addition of 1 µl of 5 N NaOH
and heating to 37°C for 10 min. One µl of 5 M HCl and 5
µl of 1 M Tris-HCl (pH 7.5) were added to neutralize the
base. Unincorporated nucleotides and salts were removed by
chromatography (Qiagen), and the cDNA was eluted in 30 µl of
dH2O. One µg of dA/dT 12–18 (Pharmacia) and 1 µg of
human Cot1 DNA (Life Technologies) were added to the probe, heat
denatured at 94°C for 5 min, combined with an equal volume of 2x
microarray hybridization solution (Amersham), and prehybridized at
50°C for 1 h. The mixture was then placed onto a microarray
slide with a coverslip and hybridized in a humid chamber at 52°C for
16 hours. The slides were washed once with 1x SSC, 0.2% SDS at room
temperature for 5 min, then twice with 0.1x SSC, 0.2% SDS at room
temperature for 10 min. After washing, the slides were rinsed in
distilled water to remove trace salts and dried.
Image Acquisition and Data Analyses.
Fluorescence intensities of the immobilized targets were measured using
a laser confocal microscope (Molecular Dynamics). Intensity data were
integrated at a pixel resolution of 10 µm using 
20 pixels per
spot, and recorded at 16 bits. Quantitative data were obtained with the
SpotFinder V 2.4 program written at the University of Washington. Local
background hybridization signals were subtracted prior to comparing
spot intensities and determining expression ratios. For each
experiment, each cDNA was represented twice on each slide, and the
experiments were performed in duplicate producing four data points per
cDNA clone per hybridization probe. Intensity ratios for each cDNA
clone, hybridized with probes derived from androgen-stimulated LNCaP
and androgen-starved LNCaP, were calculated (stimulated
intensity/starved intensity). Gene-expression levels were considered
significantly different between the two conditions if all four of the
replicate spots for a given cDNA demonstrated a ratio >2 or <0.5, and
the signal intensity was greater than 2 SD above the image background.
Cell Culture and General Methods.
DNA manipulations including transformation, plasmid preparation, gel
electrophoresis, and probe labeling, were performed according to
standard procedures (22)
. The LNCaP prostate carcinoma
cell line was cultured in RPMI 1640 supplemented with 10% FCS (Life
Technologies, Rockville, MD). Cells were transferred into RPMI 1640
with 10% CS-FCS (Life Technologies) 24 h before
androgen-regulation experiments. This medium was replaced with fresh
CS-FCS media or CS-FCS supplemented with 1 nM synthetic
androgen R1881 (NEN Life Science Products Inc.). Cells were harvested
for RNA isolation at 0-h and 72-h time points.
Northern Analysis.
Ten µg of total RNA were fractionated on 1.2% agarose denaturing
gels and transferred to nylon membranes by a capillary method
(22)
. The human multiple tissue and master blots were
obtained from Clontech. Blots were hybridized with DNA probes labeled
with [
-32P]dCTP by random priming using the Rediprime
II random primer labeling system (Amersham) according to the
manufacturer’s protocol. Filters were imaged and quantitated by using
a phosphor-capture screen and Image-quant software (Molecular
Dynamics).
cDNA Library Screening and RACE.
We screened 1,200,000 phage plaques from a human prostate 5'-stretch
cDNA library (Clontech) with the 6A4 cDNA probe representing the 3' end
of the PSDR1 cDNA. Two separate rounds of library screening
identified 16 partial-length cDNA clones. Searches of dbEST identified
seven IMAGE cDNA clones (IMAGE CloneID: 360400, 109237, 1130518,
1401718, 1337270, 1723130, 1703429) that contained sequences homologous
to PSDR1. All of the clones were sequenced and assembled
using the Sequencher software (Gene Codes, Corp.). To clone the 5' end
of the cDNA, 5'-RACE was performed on human prostate Marathon-Ready
cDNA (Clontech) using primers 6A3_RC3
(5'-GGACAGCATTTTCCTGATTTGGGGC-3') and 6A4_RC4
(5'-CAGAAGGAGGAGCAACAGCGGGAAC-3'). The RACE products were subcloned
into PCR2.1-TOPO (Invitrogen) and sequenced.
Phage plaques (1,200,000) from a human prostate 5'-STRETCH cDNA library (Clontech) were screened with PSDR1 32P-cDNA probes according to the manufacturer’s instructions. Eleven additional cDNA clones were isolated, subcloned and sequenced. RACE reactions were performed using the human prostate Marathon-ready cDNA cloning kit (Clontech) following the manufacturer’s instructions. Templates for RACE reactions were prostate Marathon-ready cDNA (Clontech) and androgen-stimulated LNCaP cDNA prepared using Marathon cDNA amplification kit (Clontech). Nested 5'-RACE reactions were performed according to the manufacturer’s instructions; first with primers 6A4RC4, 5'-CAGAAGGAGGAGCAACAGCGGGAAC-3' and AP1 (Clontech) and then a nested RACE reaction with primers 6A4RC3, 5'-GGACAGCATTTTCCTGATTTGGGGC-3' and AP2 (Clontech). The RACE products were subcloned into PCR2.1-TOPO vectors with the TOPO TA cloning kit (Invitrogen) and sequenced.
Chromosomal Localization of PSDR1 by Radiation Hybrid
Panel Mapping.
The G3 Gene bridge radiation hybrid panel (Research Genetics,
Huntsville, AL) was used to map the chromosomal localization of
PSDR1 with primers 6A4F (5'-GGGGCATTTCCTTACATTGTCTTG-3') and
6A4R (5'-CACTCCAAACAAGTGATGGGAACAC-3'). After 35 cycles of
amplification, the reaction products were separated on a 1.2% agarose
gel, and the resulting product pattern was analyzed through the
Stanford genome center web
server4
to determine the probable chromosomal location.
In Situ Hybridization.
For mRNA in situ hybridization, recombinant plasmid
pCRII-TOPO (Invitrogen), containing a 400-bp PSDR1 fragment
was linearized to generate sense and antisense digoxigenin-labeled RNA
probes. In situ hybridization was performed according to the
manufacturer’s protocol on the Ventana GenII automated instrument
(Ventana Medical Systems, Tucson, AZ). Tissue sections (5 µm) were
mounted onto Chroma plus slides (VWR Scientific), deparaffinized in a
65°C oven for 2 h followed by three 5-min soaks in xylene and
rehydrated through graded alcohol with a final rinse in 2x SSC. Before
hybridization, sections were digested with proteinase I cocktail for 12
min at 37°C, then 10 ng of either sense or antisense probe in the
hybridization buffer was applied. Programmed recipe files consisting of
buffer rinses, protease digestion, hybridization, detection, and
counterstains were optimized for the PSDR1 probe.
Digoxigenin-labeled RNA probe was added manually. Antidigoxigenin was
used as the primary antibody. The probe was denatured at 65°C, and
hybridization was carried out at 42°C for 360 min. Washes were
performed at 37°C with 2x, 1x, and 0.1x SSC. The system uses a
cocktail of antirabbit and antimouse secondary IgG-biotinylated
antibody with an indirect biotin-avidin diaminobenzidine detection
system. The sections were counterstained with hematoxylin.
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RESULTS |
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TopABSTRACTINTRODUCTIONMATERIALS AND METHODSRESULTSDISCUSSIONREFERENCES |
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. Sequence comparisons against the GenBank and dbEST databases revealed
homology only to uncharacterized partial-length ESTs (e.g.,
AA657851, IMAGE ID:1207405). Full-length cloning of the corresponding
cDNA and subsequent nucleotide and amino acid sequence comparisons
revealed significant homology to conserved motifs of the SDR family of
proteins. We have named this gene PSDR1 for Prostate
Short-chain Dehydrogenase/Reductase 1.
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. The cDNA sequence was submitted to
GenBank under the accession no. AF167438. The PSDR1 cDNA encodes a putative protein of 318 amino acids. The start codon, GAGATGG matches in a strong context to the Kozak translation initiation consensus sequences (RNNATGG, where R is a purine; Ref. 28 ). Two potential polyadenylations signals were identified at nucleotide positions 2439 and 2481. IMAGE clone 1703429 has a poly(A) stretch that uses the AATAAA polyadenylation signal at 2419, and our original cDNA clone 6A4 uses the AATAAA signals at 2481. However, we were not able to find a polyadenylation site that would produce the 900-bp band seen in testis tissue. PCR primers flanking the start and stop codons were designed, and an expected size band encompassing the entire coding region was amplified from human prostate Marathon-Ready cDNA (Clontech; data not shown).
Comparisons of the assembled cDNA sequences indicated several polymorphic sites. Five distinct single nucleotide polymorphisms were recognized between the three independent prostate tissue sources used for PSDR1 cloning. Three occur in the coding region of the PSDR1 sequence; nucleotide 379, ggc to ggg; nucleotide 916, gtg to gtc; and nucleotide 921, gtc to gcc. The first two are conserved changes, whereas the latter results in a valine to alanine amino acid substitution. Alignments of sequences in dbEST with homology to PSDR1 identified more than 20 distinct nucleotide differences in tissue sources presumably derived from different individuals.
Prostate-localized and Androgen-regulated Expression of
PSDR1.
The androgen-regulated expression of PSDR1 was confirmed by
Northern analysis using the same LNCaP RNA that was used for microarray
analysis. PhosphorImage quantitation of the Northern analysis
demonstrated a 3-fold induction of PSDR1 expression after
72 h of androgen exposure relative to 72 h of androgen
starvation (Fig. 1B)
. PSA expression increased 25-fold, and
the expression of the G3PDH loading control did not change
significantly. Interestingly, Northern analysis with
androgen-independent prostate cancer cell lines DU145 and PC3
demonstrated PSDR1 expression in both cell types (Fig. 1C)
indicating a mechanism of PSDR1 transcription
in these cells that is independent of androgen requirements.
The distribution of PSDR1 transcripts in normal human
tissues and prostate carcinoma was determined by Northern analysis and
mRNA dot blot. Of 16 adult tissues examined by Northern, a
PSDR1 message of 2.5 kb was predominantly expressed in
prostate (Fig. 1D)
. In testis, the PSDR1 probe
hybridized to an additional band at about 900 bp (Fig. 1D)
,
which could indicate cross-hybridization, alternate splicing, or
alternate usage of polyadenylation signals. The PSDR1
expression profile was confirmed using an RNA Master dot blot
(Clontech) comprised of RNA from 50 different tissues. PSDR1
expression was detected predominantly in prostate with a very low
relative level of expression in spleen, thymus, testis, ovary, small
intestine, colon, peripheral blood leukocyte, and kidney, adrenal
gland, and fetal liver (Fig. 1E)
. PSDR1
expression was at least 4-fold higher in prostate relative to any other
human tissue examined. PSDR1 expression was detected in all
of the normal and neoplastic prostate tissue samples examined. These
included three normal whole prostate tissues; three primary prostate
adenocarcinomas, androgen-dependent and androgen-independent prostate
cancer xenografts; and three prostate cancer cell lines (Fig. 1C)
.
PSDR1 Shares Homology with Members of the SDR Family.
We used the nucleotide and translated the 318-amino acid
PSDR1 sequence to search the National Center for
Biotechnology Information sequence databases by using BLAST and BEAUTY
algorithms (29)
. Partial homology was seen with several
oxidoreductases from bacteria and plant sources. To see whether the
homology was significant, we searched the protein sequence of
PSDR1 against the BLOCKS database
(30)
.5
The PSDR1 protein has three blocks that all match to the SDR
family protein signature BLOCK (BL00061; Ref. 31
) with a significant
combined E-value of 2.6e-06. The SDR family are NAD- or NADP-dependent
oxidoreductases (31)
, which include enzymes involved in
steroid metabolism such as estradiol 17-ß-dehydrogenase (also called
17-ß-hydroxysteroid dehydrogenase; EC 1.1.1.62),
15-hydroxyprostaglandin dehydrogenase (NAD+) (EC 1.1.1.141) from human
and 11 ß-HSD (EC 1.1.1.146; 11-DH; Ref. 31
). A multiple sequence
alignment of the PSDR1 protein with different members of the
human HSD family and a prokaryotic 20-ß-HSD (Streptomyces
3/20ß-HSD) is shown in Fig. 2
.
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). Sequence
alignments reveal that proteins in the SDR family exhibit residue
identities of only about 15–30%, probably because of their
early divergence and remote origin (31)
. PSDR1
shows 25% amino acid identity with other members of the SDR family.
Searches against prosite patterns
database6
revealed that PSDR1 contains two Asn-glycosylation sites at
amino acid (aa) position 174 and 198. These two sites are also
conserved among SDR family proteins (Fig. 2)
. In addition, two protein
kinase C (PKC) phosphorylation sites (aa 57 and 106), a casein kinase
II phosphorylation site (aa 57), and a 7 N-myristoylation
site are identified in the protein.
PSDR1 Genomic Organization and Promoter Sequence
Analysis.
BLAST searches with the full-length PSDR1 cDNA identified
homology with nucleotide sequence derived from a recently deposited
unannotated 197-kb chromosome 14 BAC clone, R-1012A1, sequenced by the
National Sequencing Center-Genoscope in France (GenBank accession no.
AL049779). Alignment with the PSDR1 sequence demonstrated
that this BAC contains the entire PSDR1 cDNA and allowed for
the determination of the PSDR1 genomic structure. The
PSDR1 gene comprises 7 exons and 6 introns. The sizes of
exons and introns and the exon/intron junctional sequences are listed
in Fig. 3B
. All of the intron/exon junctions conform to the 5'-gt...3'-ag
consensus (33)
except intron 2. Intron 2 has a
5'-gc...3'-ag splicing signal, a structure that has been identified
in other genes (34)
.
|
. A
sequence that has 86.7% homology (13 of 15 nucleotides) to the
consensus ARE, 5'-GGA/TACAnnnTGTTCT-3', (37)
was
identified (Fig. 3A)
. Two sequences that have 86.7% (13 of
15 nucleotides) homology to the consensus sequence of PREs
(38)
were also identified (Fig. 3A)
. An
interleukin-6 response element binding protein site, TTCCCAGAA,
(39)
was identified 281 bp 5' of the transcription
initiation site.
Chromosomal Localization of PSDR1.
The medium-resolution Stanford G3 radiation hybrid panel was used to
determine the chromosomal localization of PSDR1 using
gene-specific PCR primers 6A4F and 6A4R. Analysis of the typing results
on the Stanford Human Genome Center Radiation Hybrid Panel
server4
indicated that PSDR1 is located closest
to Stanford Human Genome Center Radiation Hybrid Panel-2558 between two
cytogenetically mapped markers D4S63 (mapped to 14q23) and D4S258
(mapped to
14q24.3).9
Therefore, PSDR1 is mapped to 14q23-24.3, consistent with
the BAC clone mapping data localizing BAC R-1012A1 to
chromosome 14q.
PSDR1 Expression in Normal and Neoplastic Prostate
Epithelium.
Normal prostate contains two major epithelial cell populations, the
luminal secretory cells and the basal cells. In situ
hybridizations were performed on sections of normal prostate by using
an antisense RNA probe specific for PSDR1 to localize its
expression. PSDR1 was expressed in both normal basal and
luminal cell populations. (Fig. 4, A and C)
. Little to no staining was seen in fibromuscular
stromal cells, endothelial cells, or infiltrating lymphocytes.
Hybridization with sense PSDR1 RNA probes showed no
background staining (Fig. 4, B and D)
. In
situ hybridizations with PSDR1 antisense and sense
probes were also performed on sections of primary prostate
adenocarcinoma obtained from radical prostatectomy specimens.
Adenocarcinoma cells were uniformly positive for PSDR1
expression (Fig. 4E)
. Hybridization with sense
PSDR1 RNA probes showed no background staining (Fig. 4F)
.
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DISCUSSION |
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-DHT and testosterone into the less potent
17-keto compounds 5-androstanedione and 4-androstenedione,
respectively (44)
. This suggests that 17HSD type 2 plays a
part in the androgen metabolic pathway, resulting in the inactivation
of testosterone and 5-DHT locally in the prostate. Enzyme expression
in the prostate could, therefore, protect cells from excessive androgen
action. Because PSDR1 shows significant homology to HSD
members of the SDR family, we hypothesize that PSDR1 is also
involved in prostate cellular steroid metabolism that may include
biosynthesis and/or degradation. The pharmacological modulation of
PSDR1 activity could thus influence prostate cellular growth
and provide a new target for prostate cancer therapy.
Numerous studies support associations between molecular variations
involving genes of the androgen metabolic pathway and the development
and progression of prostate cancer (45)
. In addition to
environmental influences, racial and international variation in
prostate cancer incidence suggests that inheritable genetic factors
such as those that influence androgen biosynthesis, activation,
transport, and metabolism are operative (45)
. In addition
to polymorphic variation in the AR itself (46)
, specific
polymorphisms in the 5-reductase type 2 (SRD5A2) gene,
the enzyme converting testosterone to the more bioactive DHT, result in
increased enzyme activity and confer up to a 7-fold increased risk for
the development of prostate cancer in African-American men
(47)
. Allelic variants in the 3-ß-HSD type II
gene, encoding one of two enzymes that initiates the inactivation of
DHT, have been identified and are currently under assessment for a role
in racial/ethnic differences in prostate carcinogenesis
(48)
. Polymorphisms in PSDR1 could influence
enzyme activity and consequently result in variations in steroid
metabolism between individuals. Our preliminary analysis of the
PSDR1 sequence from three different prostate tissue sources
identified five distinct single nucleotide polymorphisms. Three occur
in the coding region of the PSDR1 sequence, one of which
results in a valine to alanine amino acid substitution. An alignment of
sequences in dbEST with homology to PSDR1 identified more
than 20 distinct nucleotide differences in tissue sources presumably
derived from different individuals. Although some of these differences
may represent sequencing artifacts, these findings warrant a more
directed study of PSDR1 variation in different ethnic
populations and in samples of prostate carcinoma.
The expression of PSDR1 is induced by synthetic androgens in LNCaP cells. The mechanism of androgen-mediated regulation of PSDR1 expression is unknown and could involve either direct AR binding to PSDR1 promoter regions or indirect activation through the modulation of intermediary transcription factors or via posttranscriptional mechanisms. Androgens have been shown to regulate expression of other oxidoreductases. The mouse alcohol dehydrogenase ADH1, which belongs to the long-chain dehydrogenase family, is induced 10–12 fold by androgens in mouse kidney cells (49) . The induction of mouse ADH1 gene by androgens seems to be AR dependent because the ADH1 gene in Tfm mice lacking functional AR was not responsive to androgens (49) . 17-ß-HSD1, which belongs to the SDR family, is stimulated by androgen through AR-mediated mechanism (50) . We have identified a putative ARE and two putative PRE sites that demonstrate a high degree of homology to the respective consensus hormone receptor binding sites. Whether these responsive elements are functional awaits further investigation. In addition, PSDR1 transcription is not entirely mediated by androgens as demonstrated by a low level of detectable PSDR1 message in androgen-starved LNCaP cells and in the androgen-independent PC3 and DU145 prostate cancer cell lines. Studies of the PSDR1 protein may identify additional mechanisms of functional regulation.
The localized expression of PSDR1 in prostate epithelium is interesting because, to our knowledge, PSDR1 is the first member of the human SDR family that is expressed predominantly in the prostate gland. Other members of SDR family exhibit tissue-restricted patterns of expression. For example, 17-ß-HSD1 is predominantly expressed in placenta and ovary (51 , 52) ; 17-ß-HSD2 is expressed in placenta and liver (53) ; 17-ß-HSD3 is expressed in testis (42) ; 17-ß-HSD4 is primarily expressed in liver and kidney (54) ; 17-ß-HSD5 is expressed most abundantly in liver and testis (41 , 55) ; and 17-ß-HSD6 is expressed equally in liver and prostate (43) .
The identification of genes with selective expression in specific organs or cell types provides an entry point for understanding biological processes that occur uniquely within a particular tissue. Genes and their cognate proteins whose expression is specific for the prostate have greatly aided the diagnosis and treatment of prostate carcinoma. The significance of the prostate predominant expression pattern of PSDR1 remains to be determined. If the tissue expression profile of the PSDR1 protein corresponds to the transcript expression profile, then PSDR1 may represent an additional target for prostate cancer diagnostic and therapeutic interventions. Cellular or humoral immunotherapy could be designed to exploit the tissue expression differential. Our hypothesis is that PSDR1 is involved in steroid synthesis and/or degradation in normal and neoplastic prostate epithelium, and, as such, it may be a key enzyme involved in maintaining intracellular balance of steroid hormones in these cells. Additional studies including the expression of PSDR1 protein and the analysis of substrate specificity and kinetics are needed to address this question.
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ACKNOWLEDGMENTS |
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FOOTNOTES |
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1 Supported in part by the CaPCURE Foundation, by
Grant CA75173-01A1 from the National Cancer Institute (NIH) (to
P. S. N.), and by gifts from the Seattle Foundation and the Lazar
Foundation. 
2 To whom requests for reprints should be
addressed, at the Division of Human Biology, Fred Hutchinson Cancer
Research Center, Mailstop D4-100, 1100 Fairview Avenue North, Seattle,
Washington 98109-1024. E-mail: pnelson{at}fhcrc.org
3 The abbreviations used are: AR, androgen
receptor; PSA, prostate-specific antigen; ARE, androgen response
element; DHT, dihydrotestosterone; EST, expressed sequence tag; dbEST,
database of ESTs; CS-FCS, charcoal-stripped FCS; PRE, progesterone
responsive element; HSD, hydroxysteroid dehydrogenase; RACE, rapid
amplification of cDNA ends; SDR, short-chain dehydrogenase
reductase; PSDR1, prostate SDR 1; hk, human glandular
kallikrein.
4 Internet address: http://shgc.stanford.edu.
5 Internet address: http://www.blocks.fhcrc.org.
6 Internet address:
http://www.isrec.isb-sib.ch/software/PSTSCAN form.html.
7 Internet address:
http://www-hgc.lbl.gov/projects/promoter.html.
8 Internet address:
http://www.cbil.upenn.edu/tess/index.html.
9 The Genome Database: http://www.gdb.org/.
The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked advertisement in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.
10 Supported in part by the CaPCURE Foundation, by
Grant CA75173-01A1 from the National Cancer Institute (NIH) (to
P. S. N.), and by gifts from the Seattle Foundation and the Lazar
Foundation.
11 To whom requests for reprints should be
addressed, at the Division of Human Biology, Fred Hutchinson Cancer
Research Center, Mailstop D4-100, 1100 Fairview Avenue North, Seattle,
Washington 98109-1024. E-mail: pnelson{at}fhcrc.org
12 The abbreviations used are: AR, androgen
receptor; AR, androgen receptor; PSA, prostate-specific antigen; ARE,
androgen response element; DHT, dihydrotestosterone; EST, expressed
sequence tag; dbEST, database of ESTs; CS-FCS, charcoal-stripped FCS;
PRE, progesterone responsive element; HSD, hydroxysteroid
dehydrogenase; RACE, rapid amplification of cDNA ends; SDR, short-chain
dehydrogenase reductase; PSDR1, prostate SDR 1; hk, human
glandular kallikrein.
13 Internet address: www.shgc.stanford.edu.
14 Internet address: http://www.blocks.fhcrc.org.
15 Internet address:
http://www.isrec.isb-sib.ch/software/PSTSCAN form.html.
16 Internet address:
http://www-hgc.lbl.gov/projects/promoter.html.
17 Internet address:
http://www.cbil.upenn.edu/tess/index.html.
18 The Genome Database: http://www.gdb.org/.
Received 6/ 7/00. Accepted 12/13/00.
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2 macroglobulin gene.. Mol. Cell. Biol., 12: 2282-2294, 1992.-hydroxysteroid dehydrogenases from rat and human prostate.. J. Biol. Chem., 272: 15959-15966, 1997.-hydroxysteroid dehydrogenase activity. J. Biol. Chem., 268: 12964-12969, 1993.-hydroxysteroid dehydrogenase (HSD) from human prostate: demonstration of bifunctional 3/17-ß-HSD activity and cellular distribution (published erratum appears in Mol. Endocrinol., 12; 1763, 1999). Mol. Endocrinol., 11: 1971–1984, 1997.
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 J. Lapointe, C. Li, J. P. Higgins, M. van de Rijn, E. Bair, K. Montgomery, M. Ferrari, L. Egevad, W. Rayford, U. Bergerheim, et al. Gene expression profiling identifies clinically relevant subtypes of prostate cancer PNAS, January 20, 2004; 101(3): 811 - 816. [Abstract] [Full Text] [PDF]
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A. Kasus-Jacobi, J. Ou, Y. K. Bashmakov, J. M. Shelton, J. A. Richardson, J. L. Goldstein, and M. S. Brown Characterization of Mouse Short-chain Aldehyde Reductase (SCALD), an Enzyme Regulated by Sterol Regulatory Element-binding Proteins J. Biol. Chem., August 22, 2003; 278(34): 32380 - 32389. [Abstract] [Full Text] [PDF]
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F. Haeseleer, G.-F. Jang, Y. Imanishi, C. A. G. G. Driessen, M. Matsumura, P. S. Nelson, and K. Palczewski Dual-substrate Specificity Short Chain Retinol Dehydrogenases from the Vertebrate Retina J. Biol. Chem., November 15, 2002; 277(47): 45537 - 45546. [Abstract] [Full Text] [PDF]
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N. Y. Kedishvili, O. V. Chumakova, S. V. Chetyrkin, O. V. Belyaeva, E. A. Lapshina, D. W. Lin, M. Matsumura, and P. S. Nelson Evidence That the Human Gene for Prostate Short-chain Dehydrogenase/Reductase (PSDR1) Encodes a Novel Retinal Reductase (RalR1) J. Biol. Chem., August 2, 2002; 277(32): 28909 - 28915. [Abstract] [Full Text] [PDF]
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C. C. Pritchard, L. Hsu, J. Delrow, and P. S. Nelson Project normal: Defining normal variance in mouse gene expression PNAS, November 6, 2001; 98(23): 13266 - 13271. [Abstract] [Full Text] [PDF]
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