Vol.

176,

No.

April

15,

1991

1, 1991

BIOCHEMICAL

AND

BIOPHYSICAL

RESEARCH

COMMUNICATIONS Pages

HOMOLOGOUS STRUCTURES OF RUCLRAR ARD GTPase-LINKED PLASRA RECEPTOR SUGGRST ARALOGOUS MEXHANISMS OF ACTION Jouko Research Unit, University

Collagen Biochemistry,

Received

February

19,

RRRRRARE

Oikarinen

Biocenter and Oulu, Kajaanintie Finland

of

343-348

Department 52A,

of SF-90220

Medical Oulu,

1991

The molecular structures of nuclear and GTPase-linked plasma membrane receptors are compared here in the light of a recent finding suggesting that histone I31 may be an ATP/GTPase involved in transduction of the action of nuclear receptors. Considerable homology and conservation of the regions responsible for the interaction of the plasma membrane receptors with GTPases was observed in the nuclear receptors, thus suggesting analogous mechanisms of action and a common evolutionary origin for the two receptor families0 1991 zLc.3derncme**, Inc.

A large

family

of structurally

ligands

reside

exchange

in a variety

in

as transducin the

binding

of

cyclase,

Similarly,

GAP

the

synthesis

of

GTPases

of

a variety

of such

enables

dissociation

dimerization bring

about

induction which involve specifically inducible histone

of

a

change

or repression the

induction

disruption gene

certain

brought or

has

effects

for

(4,5),

been

as this

studied

in

catalyze

from

depository

about a more

are in

changes

unknown,

subtle

may

hsp90

or and

(13)

are

thought

that

to

results

in

The mechanisms although

structural

the

to

acid The ligand

receptors

(10,15-18).

binding

retinoic

interaction.

structure

genes

through

TX,

this

chromatin

nucleosome These

(16-18).

such

subsequently

to DNA, nuclear

of

in

capable,

proteins ras-p21

action

their

receptor

the

is

for

hormone,

needed

in

(15)

positioned acetylation

the

Upon binding

(14).

involved

messengers.

exert is

such

and phospholipase-O.

proteins

as steroid

1,25-dihydroxycholecalciferol

are

being

effector

of

GDP/GTP

GTPases,

signals,

as an effector

second

promoting

GoS and Goi,

with

mechanism

extracellular

These

phosphodiesterase

receptors

A ligand

by

ligand-induced

The effector

for

(l-3).

of

interacting

whose

(6-9).

nuclear

(10-12).

the

be regarded

may

so far

In general, DNA

GTP,

and act

subtypes

of

receptors

proteins

cGMP-specific

one of the detail

most

membrane

and various coupling

adenylate being

plasma

of GTP-binding

(GoT)

receptor-effector upon

the

homologous

promoter

be accompanied

by

they change

region

may

in of

by changes

a the in

(19).

0006-291X/91

343

$1.50

Vol.

176,

No.

Histone such

BIOCHEMICAL

1, 1991

Hl

is ADP,

as GDP,

(21).

Hl

areas

as compared

although

is it

is

action

of nuclear

with

It

hinge/linker

indeed

(24).

which

loosely not

to

We have

be assumed

support

of

these

suggest

that

through

that

to

this

two receptor nuclear

the

part

of

of

are

a hypoof

the

of which plasma

may

membrane

DNA and chromatin of

their

Hl

from

action.

the

the

In order

known

compared plasma

DNA

active (22,23),

forward

action to

hypothesis,

families

to

in

transduction

binding

and GTPase-linked

analogous

put

in

dissociation

least

binding

GTPase-linked

upon

cause at

its

recently

the

nucleotides

expressed

mechanism

of

COMMUNICATIONS

to chromatin

involved the

that

region

modulate

bound

as efficiently

(25),

receptors

further

may act

present

may thus

nucleosome lend

of binding

areas

as similar

nuclear

structures

capable

the

receptors

activation to

repressor

Hl may be an ATP/GTPase

be regarded

receptors.

RESEARCH

to be less

still

thesis

BIOPHYSICAL

GTP and ATP (20),

thought

that

thus

a eukaryotic

AND

molecular

here.

The results

membrane

receptors

mechanisms. METHODS

used Data

The computer programs described by Pustell and Kafatos (26-28) were and the GenebankR Genetic Sequence for protein sequence analysis, Bank to search for various sequences. RESULTS AND DISCUSSION

A

considerable

their

primary

of

structures

nuclear

membrane

degree

human

glucocorticoid

one of

the

the of

highest

homology.

similar and

higher

plots membrane regions

the

receptor

cellular

of

more

the

plasma

findings

function

molecules to

(D2R)

is

in Fig.

2. These

at

to

the

the

also

1B). ends

receptors

(see

receptors.

relate

to what

is

(TMI to ~~71

regions

to The of

fold hydropathy trans-

below), at

to

analysis the

these

present

receptors responsible

of

comprise for

and correspondingly

intracellular 344

26 % identity acid

the

D2R and GR? The polypeptide

membrane

membrane,

known

the

two display

likely

nuclear

and four

and

by Chou-Fasman (Fig.

of

(Fig.

(29)

150-amino

C-terminal

domains

coupling

so far, the

plots

membrane

domains

EC4)

GTPase

identified

D2R and GR are

plasma

transmembrane (EC1

for

in the

of various

all'GTPase-linked

plasma

GTPase-linked

receptor

between

of

of

dopamine shown

level

domain

as demonstrable only

the

responsible

hydropathy

hydrophilic

present

and

hydrophobic

structures, similar

domains

of

of

at

DNA/ligand-binding

region

of homology

regions

demonstrable

conserved

calculated

significantly

being

structure

(30)

order

How do the chains

receptor being

highly

differ

region rat

These

their

the

a putative degrees

and 38 % similarity of

is

and a C-terminal

receptors,

Alignment

identity

between

receptors

1A).

of

domains

seven anchoring

four (ICl

extrato

IC4)

Vol.

176, No.

1991

1,

A

BIOCHEMICAL

AND BIOPHYSICAL

RESEARCH COMMUNICATIONS

hGR 30

60

90

120

150 TM5i

I I

ii

l*

B

:

II

. .. .

!!! TM6

--

**

Ill II

l **

90

TM7 .!!‘i.

TM6 I

. 12c

TM7 I ‘IV

15c

: I

0

20

40

60

60

100

120

140

160

residues

2

Fiq. 1. Panel A shows the homology hash matrix of human glucocorticoid receptor (GR; residues 434 to 594) and a putative rat dopamine receptor residues 195 to 415). A part of the variable third intracellular CD& domain (IC31 of the plasma membrane receptor has been omitted in the case of D2R, since no corresponding region can be identified in GR. Panel B shows hydropathy plots of the same region in GR (-1 and D~R (-). Black bars depict the locations of transmembrane (TM) domains TM5, TM6 and TM1 in D2R, and dotted lines the regions II, III and IV responsible for interaction with a GTPase, as depicted in Fig. 3.

them

between here

to

I

to

(Fig. have

IV,

be responsible

with

the on

the

for structure

existing

(31). been

appropriate

No similar based

3)

Four

regions

demonstrated

the

interaction

GTPase

(32,331.

IC2,

by deletion of

can be deduced data,

in

although

for

IC3

and

and point

a B-adrenergic nuclear

thorough

IC4,

numbered

mutagenesis

receptor

receptors functional

such

[email protected]) as

deletion

GR and

D2R 195-IVSFYVPFIVTLLVYIKIYIVLRKRRKRVNTKR -70- NGHAKIVNPRIAKFFEIQTMPNGKTRTSL-KTMSRRKLSQQKEKKATQM GR 434-GPPPKLCLVCSDEASGCHYGVLTCGSCKvFF~VEGQHNYLCAG~DCIIDKIRRKNCPACRYRKCL~A~NLEARTK~IKGI~A DNA dim NLS

DzR

GR

-w-w --------------------LAIvL------GVFIICW--LPFFITHILNIHCDCNIPPVLYSAFTWLGYV~AVNPIIY~FNIEFRKAFMKILHC TTGvSETSENPGNKTIVPA~PQ-LTPTLVSLLEVIEPEvLY-AGYDSSVPDSTW--RI~TLNMLGGRQVIAAVKW

Fiq. 2. Alignment of D2R and GR. hatched, and w are introduced TM6 indicate the locations of TM5, regions II, III and IV responsible Badrenergic receptors the case of functional domains of GR.

IV

ligand/dim acid residues are Identical amino maximal alignment. Black bars for and TM7 in D2R, and dotted lines the for interaction with a G protein in (flAR). Double lines depict the

345

- 415 - S94/171 ~~~

Vol.

176,

No.

BIOCHEMICAL

1, 1991

extracellular

AND

BIOPHYSICAL

epace N . . . .. . . . . .

COMMUNICATIONS

ligand

..

‘.-.~..m

RESEARCH

: SAFTWLG

cytoplasm

Fiq. 3.Topological structure of GTPase-linked plasma membrane receptors such as EAR and D$. The figure has been modified from O'Dowd et al. Seven transmemhrane domains are designated TM1 to TM7 and four (29). regions shown to be involved in receptor-G protein interaction numbered I to IV. The residues of TM3, TM5 and TM7 depicted here interact with an agonist ligand.

point

mutation

responsible

for

and

121,

analyses the

of

in the

been

These three 1Y. receptor family.

of

to

reported

coupling

locate

the

polypeptide for

Interestingly,

displaying regions

the

has been

GR (34,351).

receptor-GTPase

D2R and GR,

performed

functions

structure

domain

involved between

diverse

a tertiary

DNA-binding

have

chain the

the

II,

best

III

are

also

well

conserved

and IV areas

respective-

within

can be identified

lo-

conserved

30 %, 33 % and 21 % identity

No counterpart

(see

well-conserved

regions

are

domains

the

in GR for

nuclear

region

I of

I3AR. Region function are

II

of

of

which

is

unknown.

the

first

Zn2+

lacking

The extreme

(36). a-helix

of

nuclear

well

variable

region

nuclear

receptors

finger.

Consistent

matching

the

evidently

responsible

region

activation responsible rationale

in

III of for that

the

nuclear

of

also

in part

the of

activation functionally

which by GR.

is Roth

(see

of

these

important

regions

in

the Zn2+

coded

by two region

Zn 2+

finger

is

GR (34). the

counterpart

responsible regions

for may

Consistent are

not

families.

variable

lo-12),

a GTPase by GR.

is

second

are

the

partly

DNA binding

region

second

dimerization

346

the region

receptor

sequences The

the that

recently

and the

receptors,

signal

of

two

a-helix

two latter

the

to

to a variable

(37).

a region

the

GR (34),

receptors

and this

or within

nuclear

for

of

identified

IC3 corresponds

location

transcription

been

(11,12,34),

boundary

harbors

the

of

DNA binding

this,

a nuclear

the

finger

subtypes have

between

a number

exon-exon to

Zn '+

in IC3 corresponds with

exons

of

finger part

between

separate

In addition

Some

receptors

conserved

first

to the

N-terminal

the

particularly The

D2R corresponds

conserved

thus with

the be the

between

Vol.

176,

No.

1, 1991

D2R

and

GR,

between

the

membrane the

binding

is

responsible

for

corresponding in

the

ligand

takes

place

of

a plasma

suggest

receptors

thus

mw through

function change

receptors

region for

We present

have

promote Yli-Mayry,

and the

recently the

the

removal

T.

region

Tarkka,

thought of

R.-M.

(141,

of

was

the

the

plasma

membrane they

=v ex-

nucleotide thus

may

act

Specific

target

different of

nuclear

the

receptor

ligand-binding

buried

domain

within is

by

ligands

through

change

the

present

and that

and thereby

the

present

thought

to

be

receptor.

experimental peptides

of

evidence both

to be responsible from

The

dimerization

is

for

GTPases.

receptors

while

demonstrated

responsible

families

receptors

of the

GDP-Mg2+

is

receptors,

chromatin.

conformational

obtained

C terminus

recognition

promoting

specific

on DNA,

Synthetic

regions

i.e.

Hl from

enable

a

case

membrane

origin,

receptor

membrane

no evidence in the

and GTPase-linked

Nuclear

of

activation

hypothesis.

to one of

nuclear

mechanisms,

sites

is

ligand

receptors

homology with

ligands

plasma

responsible

of

receptor

a C-terminal

GTPase-linked

homologous

degree

two

hinge

TM3, TM5 and TM1 (33).

ATP/GTPase.

to appropriate

plasma family

membrane

these

through

molecules

IC4 of

domain

the

a by

the

by nuclear

receptor

dissociation

The

ligand

a common evolutionary

activate

mechanisms.

towards

and the

analogous

selective

supposedly

residing

receptor

that

share

in an appropriate

promoting

region

variable

form

flanked There

through

nuclear

in GR are

function

a considerable a

to

(14,32).

latter

primarily

thought

any particular

the

EC4 in plasma

to

for of

the

is

residing

in GR.

families

the

highly of

conclusion, between

interaction

of

the

case

region

COMMUNICATIONS

receptors

corresponds

domains

receptor

RESEARCH

nuclear

counterpart

recognition to

while

results

variable binding

but

of

domains

responsible

receptors,

BIOPHYSICAL

region

D2R and its

in both

region

nuclear

In

This

of

domains this

here

variable

DNA and ligand IV

AND

and ligand-binding

DNA

Region

of

a highly

receptors.

between

that

BIOCHEMICAL

and J.

support

the

D2R and GR corresponding for

Hl and enable

Mannermaa

to the

GTPase

coupling

GDP/GTP exchange Oikarinen,

(N.

unpublished

results). REFERENCES D.J. and Cunnick, J.M. (1990) Cell. Signalling 2, 99-104. 1. Takemoto, 2. Gilman, A.G. (1987) Ann. Rev. Biochem. 56, 615-649. 3. Taylor, C.W. (1990) Biochem. J. 272, l-13. 4. McCormick, F. (1989) Cell 56, 5-8. Dixon, R.A.F., Schaber, M.D., Diehl, R.E., Marshall, 5. Vogel, U.S., E.M., Sigal, I.S. and Gibbs, J-B. (1988) Nature 335, M.S., Scolnick, 90-93 Cell 60, 525-528. 6. Jurnak, F., Heffron, S. and Bergmann, E. (1990) deVos, A.M., Brunger, A., Yamaizumi, Z., I. Milburn, M.V., Tong, L., Nishimura, S. and Kim, S.-H. (1890) Science 247, 939-945. 341

Vol.

176,

No.1,

BIOCHEMICALAND

1991

BIOPHYSICALFIESEARCH

COMMUNICATIONS

Pai, E.F., Krengel, U., Petsko, G.A., Goody, R.S., Kabsch, W. and Wittinghofer, A. (1990) EMBO J. 9, 2351-2359. 9. Schlichting, I., Almo, S.C., Rapp, G., Wilson, K., Petratos, K., Lentfer, A., Wittinghofer, A., Kabsch, W., Pai, E.F., Petsko, G.A. and Goody, R.S. (19901 Nature 345* 309-3t5. 10. Beato, M. (1989) Cell 56, 335-344. 11. Umesono, K. and Evans, R.M. (1989) Cell 57, 1139-1146. 12. Berg, J.M. (1989) Cell 57, 1065-1068. Danielsen, M., Ringold, G.M. and 13. Housley, P.R., Sanchez, E.R., Pratt, W.B. (1990) J. Biol. Chem. 265, 12778-12781. 14. Forman, B.M. and Samuels, H.H. (1990) Mol. Endocrinol. 4, 12938.

1301. 15. 16.

Richard-Foy, Cordingley,

H. and Hager, M.G., Riegel,

G.L. A.T.

(1987) EMBO J. 6, 2321-2328. and Hager, G.L. (1987) Cell

48,

261-

270.

32.

and Beato, M. (1990) Cell 60, 719-731. Piiia, B., Briiggemeier,U. Briiggemeier, U., Winnacker, E.-L. and Beato, M. (1990) %m, L., EMEO J. 9, 2233-2239. Bresnick, E.H., John, S., Berard# D.S., LeFebvre, P. and Hager, G.L. (1990) Proc. Natl. Acad. Sci. U.S.A. 87, 3977-3981. Mannermaa, R.-M. and Oikarinen, J. (1991) Submitted for publication. Nilsson, P., Mannermaa, R.-M., Oikarinen, J. and Grundstrom, T. (1991) Submitted for publication. Weintraub, H. (1985) Cell 42, 705-711. Nacheva, G.A., Guschin, D-Y., Prebrazhenskaya, O.V., Karpov, V.L., Ebralidse, K.K. and Mirzabekov, A.D. (1989) Cell 58, 27-36. Ericsson, C., Grossbach, U., Bjorkroth, B. and Daneholt, B. (1990) Cell 60, 73-83. Oikarinen, J., Mannermaa, R.-M., Nilsson, P. and Grundstrom, P. (1991) J. Cell. Biochem. Suppl. 15B, 249. Pustell, J. and Kafatos, F.C. (1982) Nucl. Acids Res. 10, 51-59. Pustell, J. and Kafatos, F.C. (1984) Nucl. Acids Res. 12, 643-655. Pustell, J. and Kafatos, F.C. (1986) Nucl. Acids Res, l4? 479-488. O'Dowd, B.F., Tirpak, A., Nguyen, T., Jarvie, K.R., Israel, Y., Seeman, P. and Niznik, H.B. (1990) 262, 8-12. Hollenberg, S.M., Weinberger, C., Ong, E.S., Cerelli, G., Oro, A., Lebo, R., Thompson, E.B., Rosenfeld, M.G. and Evans, R.M. (1985) Nature 318, 635-641. Lefkowitz, R.J. and Caron, M.G. (1988) J. Biol. Chem. 263, 49934996s Strader, C.D., Sigal, I.S. and Dixon, R.A.F. (1989) FASEB J. 3,

33.

Hausdorff,

17. 18. 19.

20. 21. 22. 23.

24. 25. 26. 27. 28. 29. 30.

31.

1825-1832.

W.P.,

Caron,

M.

and Lefkowitz,

R.J.

(1990)

FASEB J.

4,

2881-2889.

34. 35.

Hard, T., Kellenbach, E., Boelens, R., Maler, E.A., Dahlman, Freedman, L.P., Carlstedt-Duke, J., Yamamoto, K.R., Gustafsson, R. (1990) A. and Kaptein, Science 249, 157-160. Schwabe, J.W.R., Neuhaus, D. and Rhodes, D. (1990) Nature 348,

K., J.458-

461. 36.

37.

Segraves, Ritchie, (1990)

W.A. and Hogness, H.H., Wang, L.-H., Nucl. Acids Res. 18,

D.S. Tsai,

(1990)

6857-6862.

348

S.,

Genes

O'Malley,

Dev. 4, 204-219. B.W. and l'sai.,

M.-J.