Download Cloning and sequence analysis of the rat ventral prostate

Volume
15 Number 22
Nucleic Acids Research
1987
Cloning and sequence analysis of the rat ventral prostate glucocorticoid receptor cDNA
Chawnshang Chang, John Kokontis, Chen-Tien Chang and Shutsung Liao
The Ben May Institute and Deportment of Biochemistry and Molecular Biology,
Chicago, IL,
University of Chicago,
USA
Submitted October
16, 1987
Accession DO.Y00489
We have isolated a 1.5 kb cDNA from a rat ventral prostate XGT11 library which encodes 65% of 5*-end of glucocorticoid receptor (GR) reading frame published recently for
rat hepatoma GR cDNA (1). The prostate sequence differs from that of hepatoma in seven
nucleotides (Fig. 1, boxed). Four of these seven nucleotide changes did not alter the
deduced amino acid sequence while three changes did alter the amino acid sequence
(Table 1). Differences between the prostate and hepatoma GR cDNA sequence may
represent different allelic forms of GR or sequencing errors.
Use of this GR cDNA to measure the GR mRNA levels in different tissues produced
the following results: rat liver 100%; human prostate 31%; rat ventral prostate 29%; rat
uterus 10%. Dot blot hybridization showed that hydrocortisone injection (lOmg/day/rat) of
2 day castrated rats increased the ventral prostate GR mRNA (per unit of DNA) levels 2
fold 24 h after injection, suggesting that GR mRNA levels are regulated by glucocorticoid
in the rat ventral prostate.
OAC TCC M A G M T a n A G C T C C C C C T O f l T M A G A C G M C T C C C T C C C M T n O C T T e f i C a G e a G A S G G G A C C C T A A T G U C n T
1
ATI
Nat Asa S*r
31 TAT MA AGC
C MC CfiA
Ln Str
u ArjCl,
Cl MG ATT CTC T CAT n t » MACBCTaACAACCMTGTICMCMOTCMCMCMCMCMCMOuaCMCMCMCMCMCM
„ 52
C M CAC CM CM CM CCA 0
A TCC AM CCC CTT
TCA CTI C ATI 6
ATC CCA C M ACA • M ACA MA CTC ATG CCS
• Str L n AU Vil Sar L N S*r U t t Cly Laa T' Nat Oly Ola Thr Gla Tbr L n T i l n t t C l *
Gla Gla Gla Ola Sin Oin Pro K
C CM CTT SflC CTT
TCC TCT —
ACA
* CAC TTT
' C9C CTT CTS GM GM MC ATT OCA MC CTC
m MT CM: n c sac TAC CCA C M c
Pro Gift Gla 6 1 / Gla Laa Sly LaaSar Sarr Cly t
Thr Aip Pha Ara L n Lea Gla Gla Sar l i t A U AM Lao
AM
G n C C A C M M C a C M O M T T C A ACS TCT
111 MT
J GCA A CGC TOT GCT ACC CCB ACA GMAM CM m CCC MA ACT
1 Fro Gla AU Pro L n Sar Sar Thr Sar
Cly C n A l l Thr Pro Thr Cla Ln Clu HM Pra Lyi Thr
AM
CA CM CM CM MT CSA MA AS CM ACC
AAC OfiA OflC M T tTO A M n C TAT CCC ACA GAC CM MC
U l CAC T d OAT (
A
M
Ara
Lrt
Sar
A M Oly 0 1 / S t r Tal L n Laa Tyr Pra Thr Asp Cla S t r
Mil
AM CAT m 0A# ITT TCC GCT — T a CCA
OAC ACA M C GM ACT CCC TGB A U TCA « G CTC TTG ATA
211 ACt TTT CAC C
Pa* Asp t
Asp Thr A u Gla S t r Pro Trp
Leu Lau l i t
t Aip Lau Ala Pht S
Sar Pro
GM C C C l V ACC M T GM OAT TCT A M CCT CTT A n n A
M l OAT CM MAC 1
B GEA GM CAT
CCA n c
I Cly t t u Asp Aip Pro Fh« L M Ltn Glaa Gl;
C N Asp Thr Asa 01a Asf C n Lrs P Lau l i t Ltu
Ait Cla Asn L
BAC ACT t
ACA GM MA
T ACT GCA CAT ACAATC n A TCA AflT CCC
CMCAGTCTCOCACTSCCCCMCTCA
m cca A u Thr 1
Pro
ATT GM CTTC ACC CCC CCC CTA ATT A MCM OAfl AMCTGCSC CCA OTT TAT TIT CM GCA MC TTT TCT QGG ACA MT
301 CAT CAT nc l i t Cla L«u
* Thr Pro Cly t i l I I * L n flln Old L n Lau Cly Fro Tal Tyr C n Gla Ala S*r Pba Sar Oly Thr Asa
T CCC An TCT On CAT MTffTB A T ACC TCT OfiA C«A CM ATI TAC CAC TAT GAC ATB MT ACA GCA T a
131 ATA
MT MA ATC
A M L n Nat S
» i l ( M Thr Sar Oly l l y Ola Hat Tyr Hit Tyr A M Nat A u Thr AU Str
lit
CM
CM
GAT
CCA A n CCT OTT MT TCT GM AAC TGC MT MO TOC CM CCC T a
361 CTT
j or on m
Lao Str Sin Cla Gla Aip
_ n Pro t a l Pha Asn Til
_
___
M l CCA GMOAC MC CTO ACT
• • • 1 ^ ItA r^^w w r ^BH niv ^ M bwi
TTI OCd CCT CTG AAC
01 TJT Str Ser Pra Gly Hit Ara Pro
Civ 6 1 H Atp Scr L n Thr Str Lea Gly Ala Laa Am Fht Pro Gly Ara Str T i l Pht im- A M Oly
421 CAT CTA M t
CCA CCT OC AMCTCIK CTO CTO TOC TCC CAT CM CCT TCA GCA TCT
T i l C n ier Asa Gla A I I Str Cly C n
« l l OAT
Au
511 AAA
Ln
M t TCC CCA CCA Tto at TAT CCC AM TOTCTT CM CCT c£* ATC AAC CTT
TtC AT? ATT OAT MA
Cn l i t I la Aip LLnn l i t Ara Ara L n Am Cy» Pra Ala Cjs Arg T/r Arf L n C n Ltu t i n Ala G l / Nit A M Lm
ACA M C MA MA AAM
MMM
Thr Ln Ln Ln
Nucleotide and deduced amino acid sequences of GR. The Cl cysteine-rich region
for DNA binding is indicated by underlining. The sites at which rat prostate GR
differs from rat hepatoma GR are boxed.
TABLE 1
COHPARBON OF HAT PSOSTATE AND HEPATOMA CELL LINE OR
cDNA NUCLEOTIDE AND AMINO ACID SEQUENCES
Amino Acid
Politic™
11
98
IIS
1J7
J«0
191
MS
Codon
AmiDO t o U
Prostate
Hepatoma C«II Line
CAU
QAC"
GOT
3AC
QAC
CTG
ACT
CAA
OCC
AOT
OAT
AAC
CTA
Aarr
Prosuto
Oln
Av
dy
Ajp
A«P
Leu
Thr
Hepatorn. O i l Lin.
Ota
CHy
Ser
A^,
A^>
Leo
S«r
REFERENCES:
U
Miesfeld, R., Rusconi, S^ Goodowski, P. J., Maler, B. A., Okret, S., Wirstrom, A.,
Gustafsson, J., and Yamamoto, K. R. (1986) Cell, 46, 389-399.
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I R L Press Limited, O x f o r d , England.
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