1M

R:

Amino terminus does not have free -NH2 but is blocked (Eisen et al. 1985; Carlstedt-Duke et al.1987).

References

16P

T:

Conflict in the tamarin GR sequence either P or S

References

24R

H:

R to K hasno change on affinity or transactivation (de Lange et al., Mol.Endo.,11, 1156-1164 [1997])

References

28M

R:

Cyanogen bromide cleaves at this methionine (Carlstedt-Duke et al.1987).

References

74V

T:

Conflict in the tamarin GR sequence either A or V.

References

98G

R:

D not G (personal communication: Sandro Rusconi [Fribourg, Switzerland])

134S

M:

Is phosphorylated in steroid treated cells (Bodwell et al. 1991) but mutation to Ala has little to no effect on biological activity (Mason and Housley 1993).

References

162S

M:

Is phosphorylated in steroid treated cells (Bodwell et al. 1991) but mutation to Ala has little to no effect on biological activity (Mason and Housley 1993).

References

171T

M:

Is phosphorylated in steroid treated cells (Bodwell et al. 1991) but mutation to Ala has little to no effect on biological activity (Mason and Housley 1993).

References

224S

M:

Is phosphorylated in steroid treated cells (Bodwell et al. 1991) but mutation to Ala has little to no effect on biological activity (Mason and Housley 1993)

M:

S to A has no effect on steroid binding (Hu et al., JBC, 269, 6571 [1994]).

References

226S

R:

G not S (personal communication: M. Garabedian and K. Yamamoto [UCSF]) and (Chang, C. et al. Nucleic Acids Res 1987 Nov 25;15(22):9603)

References

232S

M:

Is phosphorylated in steroid treated cells (Bodwell et al. 1991) but mutation to Ala has little or no effect on biological activity (Mason and Housley 1993)

M:

S to A has no effect on steroid binding and azide reduces the amount of phosphorylation (Hu et al., JBC, 269, 6571 [1994]).

References

234W

R:

W to R caused4 fold decrease in transactivation of N525 (constitutively activeGR) in mammalian cells (Iniguez-Lluhi et al, JBC, 272, 4149-4156 [1997])

References

246S

M:

Is phosphorylated in steroid treated cells (Bodwell et al. 1991) but mutation to Ala has little to no effect on biological activity (Mason and Housley 1993)

M:

S to A has no effect on steroid binding and azide reduces the amount of phosphorylation (Hu et al., JBC, 269, 6571 [1994]).

References

260N

R:

D not N (personal communication: M. Garabedian and K. Yamamoto [UCSF]) and (Chang, C. et al. Nucleic Acids Res 1987 Nov 25;15(22):9603)

References

276K

R:

R not K (personal communication: M. Garabedian and K. Yamamoto [UCSF])

327S

M:

Is phosphorylated in steroid treated cells (Bodwell et al. 1991) but mutation to Ala has little to no effect on biological activity (Mason and Housley 1993).

References

345S

M:

Conflict: T not S (Chang, C. et al. Nucleic Acids Res 1987 Nov 25;15(22):9603)

References

383N

H:

N to S has no effect on dose-response curve or ability to transactivate (Karl et al. 1993)

H:

N to S has no effect (Gaitan et al., Mol. Endo., 9, 1193-1201 (1995)).

References

409F

R:

Major chymotrypsin cleavage activated RS complex after this amino acid (Carlstedt-Duke et al.1987) but no cleavage in fusion protein of protein A with the DNA-binding domain of the human receptor (Dahlman et al. 1989).

References

413Y

R:

Major chymotrypsin cleavage activated RS complex after this amino acid (Carlstedt-Duke et al.1987); only cleavage site in fusion protein of protein A with the DNA-binding domain of the human receptor (Dahlman et al. 1989).

References

435P

H:

P to R facilitated activation of agonist (triamcinolone acetonide), but not antagonist (RU 486), bound receptors (Segard-Maurel et al. 1992).

References

439L

R:

L to P has no effect on activity in vitro or in vivo (Luisi et al. 1991).

References

440C

R:

Co-ordinated with zinc (Kellenbach et al. 1991; Luisi et al. 1991);

R:

C to A does not eliminate zinc binding but does dramatically reduce (<10%) DNA binding (Archer et al.1990).

H: C to G may change effect of receptor on IL-1alpha induction from repression to induction (Ray et al. 1991);

H:

C to Y eliminates transactivation without affecting steroid binding activity and, in combination with L771F, is responsible for the activation labile mutant (Powers et al. 1993).

References

443C

R:

Co-ordinated with zinc (Kellenbach et al. 1991; Luisi et al. 1991).

References

444S

H:

S to G caused 2 fold increased transactivation but eliminated ability to suppress AP-1 induction (Heck et al., EMBO, 13, 4087-4095 (1994)).

References

445S

H:

D to G may increase the fold induction by Dex (Guido et al., Mol. Endo. 10:1178-1190 (1996)).

References

449G

R:

449-455 - some point mutants in 407/556 had little effect on biological activity (Zandi et al. 1993).

M:

V to G has wild type properties (Kasai 1990).

References

450C

H:

C to G gave receptor with "40% transactivation but "10% DNA binding activity of wild type receptor (Hollenberg and Evans 1988).

References

451H

R:

451-462 - some point mutants (depends on mutant AA) eliminate or reduce biological activity in 407/556 (Zandi et al. 1993);

R:

H451N/S459G double mutation in truncated receptor of 407-556 has increased affinity for DNA and transactivation activity (Zandi et al. 1993).

References

452Y

R:

Part of interchain hydrophobic interactions holding 3° structure of DNA binding domain together (Luisi et al. 1991).

References

455L

H:

L to V caused 2 fold increased transactivation but eliminated ability to suppress AP-1 induction; double mutant (S444G/L455V) caused 2 fold increased activation and 50% reduction of suppression (Heck et al., EMBO 13:4087-4095 (1994)).

References

457C

R:

co-ordinated with zinc (Kellenbach et al., FEBS Letters, 291, 367-370 [1991]; Luisi et al., Nature, 352, 497-505 [1991]).

References

458G

H:

involved in specificity of DNA binding, along with other AAs (Zilliacus et al., JBC, 1992). No effect on biological activity (Umesono and Evans, Cell, 57, 1139 [1989])

H:

all receptors from saturation mutagenesis (truncated GR in yeast) were active with GRE or ERE; main function of residue was to inhibit binding to non-cognate sites (Zilliacus et al., PNAS, 91, 4175-4179 (1994))

References

459S

H:

Involved in specificity of DNA binding, along with other AAs (Zilliacus et al. 1992b).

R:

H451N/S459G double mutation in truncated receptor of 407-556 has increased affinity for DNA and transactivation activity (Zandi et al. 1993)

R:

S to A gives receptor identical to P493R that has normal DNA binding, poor transactivation in eukaryotic cells but good activation in yeast, and 10 fold increased affinity for non-specific DNA (Lefstin et al., Genes Devel.,8, 2842-2856 (1994))

H:

all receptors from saturation mutagenesis (truncated GR in yeast) were active with GRE or ERE; main function of residue was to inhibit binding to non-cognate sites (Zilliacus et al., PNAS, 91, 4175-4179 (1994))

References

460C

R:

Co-ordinated with zinc (Kellenbach et al. 1991; Luisi et al. 1991).

References

461K

R:

makes base contacts in binding to specific DNA sites (Luisi et al., Nature, 352, 497-505 [1991]).

R:

K to A has wild type non-specific DNA binding affinity but <1% transactivation (versus ~25% activity in F9 cells [Starr et al., Genes and Development 10:1271-1283 {1996}] while K to E caused 10 fold decrease in non-specific DNA binding affinity (Lefstin et al., Genes Devel. 8:2842-2856 (1994)).

H:

K to G causes GR to induce, as opposed to repress, AP-1 regulated gene expression (Lucibello et al., EMBO, 9, 2827-2834 [1990]; Yang-Yen et al., Cell, 62, 1205-1215 [1990]) and DNA binding is retained but trans-activation is lost (Hollenberg & Evans, Cell, 55, 899-906 [1988]).

H:

K to G afforded wild type, or almost no, induction by Dexamethasone depending on the GRE used (TAT or MMTV; Guido et al., Mol. Endo. 10:1178-1190 (1996)).

R:

K to A caused induction instead of repression from GRE and nGRE (Starr et al., Genes and Development, 10, 1271-1283 (1996)).

References

462V

H:

involved in specificity of DNA binding, along with other AAs (Zilliacus et al., JBC, 1992; R: Luisi et al., Nature, 352, 497-505 [1991])

R:

V to E caused 10 fold decrease in non-specific DNA binding affinity (Lefstinet al., Genes Devel.,8, 2842-2856 (1994)) and ~60% reduction of transactivation (Starr et al., Genes and Development, 10, 1271-1283 (1996)).

References

463F

R:

part of interchain hydrophobic interactions holding 3° structure of DNA binding domain together (Luisi et al., Nature, 352, 497-505 [1991]).

References

464F

R:

part of interchain hydrophobic interactions holding 3° structure of DNA binding domain together (Luisi et al., Nature, 352, 497-505 [1991]).

References

465K

H:

K to G gave receptor with "40% transactivation but "10% DNA binding activity of wild type receptor (Hollenberg & Evans, Cell, 55, 899-906 [1988]).

References

466R

R:

makes base contacts in binding to specific DNA sites. R to K or G is inactive in vivo (Luisi et al., Nature, 352, 497-505 [1991])

R:

R to A caused 10 fold decrease in non-specific DNA binding affinity (Lefstinet al., Genes Devel.,8, 2842-2856 (1994)) and almost no transactivation of a simple GRE but still is capable of repression (Starr et al., Genes and Development, 10, 1271-1283 (1996)).

References

468V

R:

468-472 - can tolerate insertion of tandem repeats of 3-9bp w/o loss of biological activity and of 23bp with partial loss (Zandi et al., 1993).

References

470G

M:

Insertion of Arg causes a 2 fold decrease in transcriptional activation (Kasai, 1990).

References

Insert after 470+

T:

Conflict in the Tamarin sequence; either no amino acid or R.

References

474Y

R:

part of interchain hydrophobic interactions holding 3° structure of DNA binding domain together (Luisi et al., Nature, 352, 497-505 [1991]).

H:

Y to G gave receptor with "40% transactivation but "10% DNA binding activity of wild type receptor (Hollenberg & Evans, Cell, 55, 899-906 [1988]).

References

476C

R:

co-ordinated with zinc (Kellenbach et al., FEBS Letters, 291, 367-370 [1991]; Luisi et al., Nature, 352, 497-505 [1991]).

References

477A

R:

A to T prevents DNA binding and transactivation from a gene regulated by a single, but not a multiple, copy of GRE but does not affect repression (personal communication: Peter Herrilich [Karlsruhe, Germany])

R:

A to T in a triple mutant ((A477T/R479D/D481C reduced transactivationby greater than 90% but had no effect on the ability to repress NF-kB activity (Linden et al., JBC, 272, 21467-21472 (1997))

H:

A to T eliminated DNA binding and transactivation but not repression (Heck et al., EMBO 13:, 4087-4095 (1994)).

References

479R

H:

thought to form inter-receptor salt bridge with D481 and thus play a major role in receptor dimerization (Dahlman-Wright et al., JSBMB, 45, 239-250 [1993]), although mutation to G has no major effect (Hollenberg & Evans, Cell, 55, 899-906 [1988])

R:

R479A/N491A abolished cooperative DNA binding in vitro and transactivation in vivo (Lefstin et al., Genes Devel.,8, 2842-2856 (1994)) without disturbing repression (Starr et al., Genes and Development 10:1271-1283 (1996))

R:

R to D decreased induction from a single GRE but increased synergism from 2 or 3 tandem GREs (Liu et al., Mol. Endo., 10, 1399-1406 (1996)).

References

R:

R to D in a triple mutant ((A477T/R479D/D481C reduced transactivationby greater than 90% but had no effect on the ability to repress NF-kB activity (Linden et al., JBC, 272, 21467-21472 (1997))

481D

R:

D to R causes greatly decreased activation in N-terminal deletion of Pearce and Yamamoto [Science, 259, 1161 (1993)] (Liu et al., PNAS 92:12480-12484 (1995))

R:

D to R decreased induction from a single GRE but increased synergism from 2 or 3 tandem GREs (Liu et al., Mol. Endo., 10, 1399-1406 (1996))

R:

D to C in a triple mutant ((A477T/R479D/D481C reduced transactivationby greater than 90% but had no effect on the ability to repress NF-kB activity (Linden et al., JBC, 272, 21467-21472 (1997))

H:

thought to form inter-receptor salt bridge with R479 and thus play a major role in receptor dimerization (Dahlman-Wright et al., JSBMB, 45, 239-250 [1993]), although mutation to G has no major effect (Hollenberg & Evans, Cell, 55, 899-906 [1988]).

References

482C

R:

co-ordinated with zinc (Kellenbach et al., FEBS Letters, 291, 367-370 [1991]; Luisi et al., Nature, 352, 497-505 [1991]).

References

484I

R:

most non-conservative point mutants eliminate both DNA binding and transactivation (Lanz et al., Steroids, 1994).

References

486K

H:

K to G gave receptor with "40% transactivation but "10% DNA binding activity of wild type receptor (Hollenberg & Evans, Cell, 55, 899-906 [1988]).

References

488R

R:

R to Q caused normal DNA binding and fair activity in CV-1 cells but <3% transcriptional activation in yeast (Schena et al., Genes & Dev., 3, 1590 [1989])

R:

R to A abolished cooperative DNA binding in vitro and transactivation in vivo (Lefstinet al., Genes Devel.,8, 2842-2856 (1994)).

R:

R to Q reduced transactivation , and repression of NF-kB activity, by 80% (Linden et al., JBC, 272, 21467-21472 (1997))

References

489R

R:

R to K caused normal DNA binding and good activity in CV-1 cells but <10% transcriptional activation, and temperature sensitive, in yeast (Schena et al., Genes & Dev., 3, 1590 [1989]).

References

490K

R:

K to E reduced transactivation , and repression of NF-kB activity, by 90% (Linden et al., JBC, 272, 21467-21472 (1997))

References

491N

R:

N to S caused normal DNA binding and good activity in yeast but <1% transcriptional activation in CV-1 cells (Schena et al., Genes & Dev., 3, 1590 [1989])

R:

N to A had essentially wild type activity for transactivation, and repression of NF-kB activity (Linden et al., JBC, 272, 21467-21472 (1997))

R:

R479A/N491A abolished cooperative DNA binding in vitro and transactivation in vivo (Lefstin et al., Genes Devel.,8, 2842-2856 (1994)) without disturbing repression (Starr et al., Genes and Development 10:1271-1283 (1996)).

References

492C

R:

co-ordinated with zinc (Kellenbach et al., FEBS Letters, 291, 367-370 [1991]; Luisi et al., Nature, 352, 497-505 [1991])

H:

C to S may cause incorrect folding of DNA binding domain (Zilliacus et al., JSBMB, 1992).

References

493P

R:

P493R, A494S double mutant caused normal DNA binding but poor transcriptional activation (Godowski et al.,(UCLA Symposium on Molecular and Cellular Biology. 95), Alan R. Liss, Inc., pp. 197-210 (1989)

R:

P to R single mutant completely reproduced the normal DNA binding and poor transactivation in eukaryotic cells, but good activation in yeast, of the P493R/A494S double mutant LS7 and is identical to S459A, which has 10 fold increased affinity for non-specific DNA (Lefstin et al., Genes Devel.,8, 2842-2856 (1994)).

References

495C

R:

co-ordinated with zinc (Kellenbach et al., FEBS Letters, 291, 367-370 [1991]; Luisi et al., Nature, 352, 497-505 [1991])

H:

C to S may cause incorrect folding of DNA binding domain (Zilliacus et al., JSBMB, 1992);

H:

double mutant (C495W/R498Q) eliminates activation and repression (Heck et al., EMBO 13:, 4087-4095 (1994)).

References

496R

M:

R to H is biologically inactive and has greatly reduced nuclear translocation (like nt-) (Danielsen et al.1986; Danielsen et al.1987);

H:

R to S has no effect on transactivation or repression (personal communication: Peter Herrilich [Karlsruhe, Germany])

H:

R to S caused 2 fold increased transactivation and 50% decrease in repression (Heck et al., EMBO, 13, 4087-4095 (1994)).

References

497Y

R:

Part of interchain hydrophobic interactions holding 3° structure of DNA binding domain together (Luisi et al. 1991);

H:

Y497L/R498G double mutation causes 25-100% increase in transactivation while eliminating repression (personal communication: Peter Herrilich [Karlsruhe, Germany])

H:

double mutant (Y497L/R498G) has little effect on activation but eliminates repression (Heck et al., EMBO, 13, 4087-4095 (1994)).

References

498R

H:

C495S/R498Q double mutation eliminates transactivation and repression while Y497L/R498G double mutation causes 25-100% increase in transactivation while eliminating repression (personal communication: Peter Herrilich [Karlsruhe, Germany])

H:

R to G caused slight decrease in activation and repression; see also C495 and Y497 (Heck et al., EMBO, 13, 4087-4095 (1994)).

References

500C

R:

C to S/A/M has little or no effect (Severne et al. 1988) but many others give inactive GR;

R:

C to R is biologically inactive (Schena et al. 1989); only C in 481/777 fragment of E. coli overexpressed hGR not labeled by Dex-Mes (Ohara-Nemoto et al. 1990).

References

505M

R:

M to L or C has little effect but others eliminate activity (personal communication: Sandro Rusconi [Fribourg, Switzerland])

H:

M to G gave receptor with 10% transactivation but only 1% DNA binding of wild type receptor (Hollenberg and Evans 1988).

References

517K

R:

Trypsin cuts activated RS complex after this amino acid (Carlstedt-Duke et al.1987).

References

522A

X:

Confict in Xenopus sequence S or P (P49844)

524A

X:

Confict in Xenopus sequence T or A (P49844)

526V

X:

Confict in Xenopus sequence T or A (P49844)

531S

X:

Confict in Xenopus sequence P or N (P49844)

Insert after 534+

T:

Added A in Tamarin is one of 4 mutations in steroid binding domain proposed to be responsible for 10 fold lower affinity (Brandon et al. 1991).

References

536K

R:

Trypsin is thought to cleave after this site in steroid-free receptors to form 16K core binding fragment (Simons et al. 1989).

References

538I

X:

Confict in Xenopus sequence L or M (P49844)

550L

R:

550 or 551 is the amino terminus of the steroid binding domain (Xu et al., JBC, 271, 21430-21438 (1996)); lack of binding by human mutant lacking amino acids 489-532 (Cadepond et al., J. Biol. Chem. 266: 5834-5841 (1991)), =507-550 in rat, argues that 550 is amino terminus.

References

551V

R:

:550 or 551 is the amino terminus of the steroid binding domain (Xu et al., JBC, 271, 21430-21438 (1996)).

References

553L

M:

L553,545G eliminated transcriptional activity (personal communication: Michael Stallcup [USC, Calif.])

M:

L553G/L554G eliminated Dex biological activity and affinity labeling by Dex-Mes (Milhon et al., JSBMB, 51, 11-19 (1994)).

References

554L

M:

L553G/L554G eliminated Dex biological activity and affinity labeling by Dex-Mes (Milhon et al., JSBMB, 51, 11-19 (1994)).

References

555E

M:

E to A requires 1.7 fold higher Dex concentrations for biological activity (Milhon et al., JSBMB, 51, 11-19 (1994)).

References

556V

M:

V to G requires 7 fold higher Dex concentrations for biological activity (Milhon et al., JSBMB, 51, 11-19 (1994)).

References

557I

M:

V to G requires 4-100 fold higher steroid concs. for biological activity (personal communication: Michael Stallcup [USC, Calif.])

References

558E

M:

E to G eliminates steroid binding activity (Danielsen et al.1986).

References

559P

M:

P to A requires "100 fold higher steroid concs. for biological activity, presumably due to decreased steroid binding affinity (Byravan et al. 1991).

References

560E

M:

E to A requires,6 fold higher Dex concentrations for biological activity (Milhonet al., Mol. Endo.,11, 1795-1805 (1997))

References

561V

M:

V to G requires 23 fold higher Dex concentrations for biological activity (Milhon et al., JSBMB, 51, 11-19 (1994)).

References

562L

M:

L to A requires13 fold higher Dex concentrations for biological activity (Milhonet al., Mol. Endo.,11, 1795-1805 (1997)).

References

563Y

X:

Confict in Xenopus sequence Y or F (P49844)

567D

M:

D to A requires 18 fold higher Dex concentrations for biological activity (Milhon et al., Mol. Endo. 11:1795-1805 (1997))

568S

T:

Conflict in the tamarin sequence either T or S

M:

S to A has wild type biological activity (Milhon et al., Mol. Endo. 11:1795-1805 (1997))

References

570V

X:

Confict in Xenopus sequence I or M (P49844)

572D

M:

D to G givesreceptor that was not stably expressed in Cos-7 cells (Milhonet al., Mol. Endo.,11, 1795-1805 (1997))

573S

M:

S to A requires9 fold higher Dex concentrations for biological activity Milhonet al., Mol. Endo.,11, 1795-1805 (1997))

577I

H:

I to N caused loss of Dexamethasone binding and gave a dominant negative receptor (Karl et al., Proc. Assoc. Amer. Phys., 108, 296-307 (1996))

581L

R:

L to P causes no activity in yeast or COS-7 cells (Garabedian and Yamamoto 1992);

M:

L to F requires 4-100 fold higher steroid concs. for biological activity [M] (personal communication: Michael Stallcup [USC, Calif.]);

M:

L to F causes 5-fold decreased affinity and 12 fold higher Dex concentrations for biological activity with some loss of steroid binding activity (Lee et al., Mol. Endo., 9, 826-837 (1995)

H:

residue is within 4.5 Å of Dex ligand in model of steroid binding domain (Wurtz et al. 1996).

References

582N

H:

Residue is within 4.5 Å of Dex ligand in model of steroid binding domain (Wurtz et al. 1996).

References

583M

R:

Cyanogen bromide cleaves at this methionine (Carlstedt-Duke et al.1987)

H:

M to R may cause 1000 fold increased affinity for Dex, but only 50 fold increased potency, with 5 fold increased maximal transactivation (Warriarer al., JBC, 269, 29010-29015 (1994)).

References

584L

R:

L to S causes no activity in yeast or COS-7 cells and decreased affinity for Dex in COS-7 cells (Garabedian and Yamamoto 1992);

H:

residue is within 4.5 Å of Dex ligand in model of steroid binding domain (Wurtz et al. 1996).

References

585G

H:

Residue is within 4.5 Å of Dex ligand in model of steroid binding domain (Wurtz et al. 1996)

H:

G to C eliminated steroid binding and transactivation (Benhamou et al., Science 255:206-299 (1992));

H:

G to A destroyed steroid binding and transactivation (Warriar er al., JBC, 269, 29010-29015 (1994)).

References

588Q

H:

Residue is within 4.5 Å of Dex ligand in model of steroid binding domain (Wurtz et al. 1996).

References

591A

H:

A to Q may cause 1000 fold increased affinity for Dex, but only 7 fold increased potency, with 5 fold increased maximal transactivation (Warriarer al., JBC, 269, 29010-29015 (1994)).

References

600L

R:

P not L (personal communication: M. Garabedian and K. Yamamoto [UCSF])

602L

R:

F not L (personal communication: M. Garabedian and K. Yamamoto [UCSF])

618W

M:

W to A has no effect on biological activity (personal communication: Michael Stallcup [USC, Calif.]);

H:

residue is within 4.5 Å of Dex ligand in model of steroid binding domain (Wurtz et al. 1996).

References

619M

H:

M to L causes 2-3 fold reduced affinity for Dex (personal communication: Jan Carlstedt-Duke [Huddinge, Sweden])

620F

R:

F to S causes increased affinity for Dex and triamcinolone acetonide in yeast, but not in CV-1 cells, and thus suggests some non-receptor factor in steroid binding (Garabedian and Yamamoto 1992).

References

622M

R:

M to P causes reduced transcriptional activity in yeast (Garabedian and Yamamoto 1992);

M:

M to L, C, or S has no effect on biological activity (personal communication: Michael Stallcup [USC, Calif.]);

R+H: photo-labeled by triamcinolone acetonide(R, H) (Carlstedt-Duke et al. 1988; Stromstedt et al. 1990) and R5020 [R] (Stromstedt et al. 1990); cyanogen bromide cleaves at this methionine (Carlstedt-Duke et al.1987);

H:

M to L has no significant effect on affinity (personal communication: Jan Carlstedt-Duke [Huddinge, Sweden]);

H:

residue is within 4.5 Å of Dex ligand in model of steroid binding domain (Wurtz et al. 1996).

References

626L

H:

Residue is within 4.5 Å of Dex ligand in model of steroid binding domain (Wurtz et al. 1996)

References

628W

M:

W to A requires "100 fold higher steroid concs. for biological activity (personal communication: Michael Stallcup [USC, Calif.])

629R

M:

R to A eliminates transcriptional activity (personal communication: Michael Stallcup [USC, Calif.]);

H:

residue is within 4.5 Å of Dex ligand in model of steroid binding domain (Wurtz et al. 1996).

References

634S

S to A in marmoset is one of 4 mutations in steroid binding domain proposed to be responsible for 10 fold lower affinity (Brandon et al. 1991).

References

636G

A to S in marmoset is one of 4 mutations in steroid binding domain proposed to be responsible for 10 fold lower affinity (Brandon et al. 1991).

References

640C

R:

Forms an intramolecular disulfide with C656 or C661 (Chakraborti et al. 1992);

R:

C to S causes 3 fold loss in binding affinity (Chakraborti et al. 1991);

M:

C to A has no effect on biological activity (Chen and Stallcup 1994)

H:

residue is within 4.5 Å of Dex ligand in model of steroid binding domain (Wurtz et al. 1996).

H:

labeled by Dex-Mes in 481/777 of hGR overexpressed in E. coli (Ohara-Nemoto et al. 1990).

References

641F

M:

F to A requires 4-100 fold higher steroid concs. for biological activity (personal communication: Michael Stallcup [USC, Calif.]);

H:

residue is within 4.5 Å of Dex ligand in model of steroid binding domain (Wurtz et al. 1996).

References

643P

M:

P to A requires "100 fold higher steroid concs. for biological activity (personal communication: Michael Stallcup [USC, Calif.])

656C

R:

Covalently labeled by Dex-Mes (Simons et al.1987; Carlstedt-Duke et al. 1988);

M:

Covalently labeled by Dex-Mes (Smith et al. 1988);

H:

Covalently labeled by Dex-Mes (Stromstedt et al. 1990);

R:

Forms an intramolecular disulfide with C661 or C640 (Chakraborti et al. 1992);

R:

forms a specific complex with arsenite (Lopez et al. 1990; Chakraborti et al. 1992);

H:

residue is within 4.5 Å of Dex ligand in model of steroid binding domain (Wurtz et al. 1996).

R:

C to G or S yields "super" receptor (Chakraborti et al. 1991);

R:

C to Y has no effect on affinity of receptors expressed in COS-7 cells (Chakraborti and Simons, unpublished results) but reduces transcriptional activity in yeast (Garabedian and Yamamoto 1992);

H:

C to W has no effect on transcriptional activity (Keightley and Fuller 1994);

H:

C to S has no effect on affinity (personal communication: Jan Carlstedt-Duke [Huddinge, Sweden])

H:

C to S causes about 15 fold higher affinity (Yu et al., Biochem. 34:14163-14173 (1995).

M:

C to S has no effect on biological activity (Chen and Stallcup, JBC 269:7914-7918 (1994));

References

657M

H:

Residue is within 4.5 Å of Dex ligand in model of steroid binding domain (Wurtz et al. 1996).

References

659D

R:

D to G causes no activity in yeast or COS-7 cells and decreased affinity for Dex in COS-7 cells (Garabedian and Yamamoto 1992);

H:

D to V reducedaffinity 3 fold and transactivation 25 fold but had no effecton repression (de Lange et al., Mol. Endo.,11, 1156-1164[1997])

H:

D to V causes 3 fold decreased steroid affinity and 7 fold reduced glucocorticoid sensitivity (Hurley et al. 1991).

References

661C

R:

Forms an intramolecular disulfide with C656 or C640 ((Chakraborti et al. 1992);

R:

forms a specific complex with arsenite (Lopez et al. 1990; Chakraborti et al. 1992);

R:

C to S causes 4 fold loss in binding affinity (Chakraborti et al. 1991);

R:

C to R is transcriptionally active in yeast only with deacylcortivazol (Garabedian and Yamamoto 1992);

M:

C to G requires 2-3 fold higher steroid concentrations for biological activity (Chen and Stallcup 1994).

H:

labeled by Dex-Mes in 481/777 of hGR overexpressed in E. coli (Ohara-Nemoto et al. 1990).

H:

C to S has wild type affinity in cell-free assay (Yu et al., Biochem. 34:14163-14173 (1995)).

References

664M

R:

Cyanogen bromide cleaves at this methionine (Carlstedt-Duke et al.1987).

References

671L

R:

L to S causes reduced transcript. activity in yeast or COS-7 cells and decreased affinity for Dex in COS-7 cells (Garabedian and Yamamoto 1992).

References

673R

X:

Confict in Xenopus sequence S or R (P49844)

R:

Trypsin is thought to cleave after this site in steroid-free receptors to form 16K core binding fragment (Simons et al. 1989).

References

682L

M:

L to F requires 4-100 fold higher steroid concs. for biological activity [M] (personal communication: Michael Stallcup [USC, Calif.])

M:

L to F causes activation labile phenotype with no loss in Dex affinity (but 200 fold higher Dex concentrations are required for biological activity) and changes in relative affinity only for DAC (Lee et al., Mol. Endo., 9, 826-837 (1995)).

References

683C

M:

C to S or A requires 4-100 fold higher steroid concs. for biological activity (Chen and Stallcup 1994)

M:

C683S/C754S double mutant caused a 300 fold increase in the Dex concentration required for induction (Chen and Stallcup, JBC 269:7914-7918 (1994));

H:

both C to S and C683S/M684L double mutant have no effect on affinity (Yu et al., Biochem., 34, 14163-14173 (1995))

H:

labeled by Dex-Mes in 481/777 of hGR overexpressed in E. coli (Ohara-Nemoto et al. 1990).

H:

C to S has no effect on affinity (personal communication: Jan Carlstedt-Duke [Huddinge, Sweden])

References

684M

R:

Cyanogen bromide cleaves at this methionine (Carlstedt-Duke et al.1987);

M:

M to I requires 4-100 fold higher steroid concs. for biological activity (personal communication: Michael Stallcup [USC, Calif.])

M:

M to I causes about 3 fold lower Dex affinity (10 fold higher Dex concentrations are required for biological activity) and some loss of binding specificity (Lee et al., Mol. Endo., 9, 826-837 (1995));

H:

C683S/M684L double mutant has no effect on affinity (Yu et al., Biochem. 34:14163-14173 (1995)).

References

706E

R:

E to K causes no activity in yeast or COS-7 cells and decreased affinity for Dex in COS-7 cells (Garabedian and Yamamoto 1992).

References

715L

R:

L to V has no effect in yeast (Garabedian and Yamamoto 1992).

References

732R

R:

R to Q caused about a 5 fold a right shift in the Dexamethasone dose-response curve in full length receptor and eliminated steroid binding in context of del 550-795 (Xu and Simons, personal communication).

747V

H:

V to I in patient with 1° cortisol resistance causes 2-fold decreased affinity and 4-fold decreased EC50 (Malchoff et al. 1993).

H:

V to I reducedaffinity by 2 fold and transactivation 10 fold but only marginallyreduced repression (de Lange et al., Mol. Endo.,11, 1156-1164 [1997])

References

754C

R:

Photolabeled by triamcinolone acetonide (Carlstedt-Duke et al. 1988) and R5020 (Stromstedt et al. 1990);

M:

C to G requires >100 fold higher steroid concs. for biological activity, apparently due to decreased protein stablity and degradation to a 68 kDa species (no 98 kDa seen) (Byravan et al. 1991) and C to S requires 4-100 fold more steroid for biological activity (Chen and Stallcup 1994)

M:

C683S/C754S double mutant caused a 300 fold increase in the Dex concentration required for induction (Chen and Stallcup, JBC 269:7914-7918 (1994))

H:

labeled by Dex-Mes in 481/777 of hGR overexpressed in E. coli (Ohara-Nemoto et al. 1990);

H:

residue is within 4.5 Å of Dex ligand in model of steroid binding domain (Wurtz et al. 1996).

H:

saturation mutagenesis revealed that only 3 changes ( to A, S. or T) are compatible with binding, with S and T mutations differently affecting the binding of triamcinolone acetonide and cortisol (Lind et al., Mol. Endo., 10, 1358-1370 (1996)).

References

757T

H:

Residue is within 4.5 Å of Dex ligand in model of steroid binding domain (Wurtz et al. 1996).

References

579L

X:

Confict in Xenopus sequence L or M (P49844)

761K

H:

Residue is within 4.5 Å of Dex ligand in model of steroid binding domain (Wurtz et al. 1996).

References

762T

R:

T to I causes reduced transcript. activity in yeast (Garabedian and Yamamoto 1992).

References

765I

H:

I to T reduced affinity for Dex or cortisol by less than or equal to 2 fold but caused a 100 fold right shift in the dose response curve and is essentially inactive with natural glucocorticoids (Roux et al., Mol. Endo. 10:1214-1226 (1996)).

References

767F

H:

Residue is within 4.5 Å of Dex ligand in model of steroid binding domain (Wurtz et al. 1996).

References

770M

M:

Cyanogen bromide cleaves at this methionine (Carlstedt-Duke et al.1987).

M:

Double mutation to A (770/771) has no major affect on steroid binding but eliminates transcriptional activation (Danielian et al. 1992). However, in rat receptors, this mutation has either a three fold lower affinity for steroid (Schmitt and Stunnenberg1993) or no affinity at all (Lanz et al. 1994)

R:

M770A/L771A caused loss of binding of Dex, but not RU 486, which displayed partial agonist activity of wild type receptor (Lanz and Rusconi, Endo., 135, 2183-2195 (1994)).

References

771L

R:

M770A/L771A caused loss of binding of Dex, but not RU 486, which displayed partial agonist activity of wild type receptor (Lanz and Rusconi, Endo., 135, 2183-2195 (1994))

M:

Double mutation to A (770/771) has no major affect on steroid binding but eliminates transcriptional activation (Danielian et al. 1992). However, in rat receptors, this mutation has either a three fold lower affinity for steroid (Schmitt and Stunnenberg1993) or no affinity at all (Lanz et al. 1994).

H:

L to F gives rise to activation labile or r- phenotype, depending on the cellular environment (Ashraf and Thompson 1993); this same mutation gives receptors with unstable binding at 37°C but not at 0°C and, in combination with C440Y, will also give an activation labile phenotype (Powers et al. 1993).

H:

Residue is within 4.5 Å of Dex ligand in model of steroid binding domain (Wurtz et al. 1996).

References

773E

M:

E to A does not affect steroid binding but decreases transcriptional activation by Å40% (Danielian et al. 1992).

References

774I

M:

I774A/I775A double mutant has no major affect on steroid binding but eliminates transcriptional activation (Danielian et al. 1992);

R:

however, Lanz et al. found that these mutations did not bind steroid (Lanz et al. 1994).

R:

I774A/I775A caused loss of binding of Dex, but not RU 486, which displayed partial agonist activity of wild type receptor (Lanz and Rusconi, Endo., 135, 2183-2195 (1994)).

References

775I

M:

I774A/I775A double mutant has no major affect on steroid binding but eliminates transcriptional activation (Danielian et al. 1992);

R:

however, Lanz et al. found that these mutations did not bind steroid (Lanz et al. 1994).

R:

I774A/I775A caused loss of binding of Dex, but not RU 486, which displayed partial agonist activity of wild type receptor (Lanz and Rusconi, Endo., 135, 2183-2195 (1994))

H:

Residue is within 4.5 Å of Dex ligand in model of steroid binding domain (Wurtz et al. 1996).

References

779I

T:

Conflict in the Tamarin sequence either L or I.

References

780P

R:

P780/K781 double deletion has no effect on activity with either Dex or RU486 (Lanz and Rusconi, Endo., 135, 2183-2195 (1994)).

References

781K

R:

P780/K781 double deletion has no effect on activity with either Dex or RU486 (Lanz and Rusconi, Endo., 135, 2183-2195 (1994)).

References

782Y

M:

Y to N needs 3 to 4 fold more Dex for equal biological activity, presumably due to 3-4 fold lower affinity (Danielsen et al. 1986)

M:

Y to N caused 2-10 fold right shift in dose-response curves of numerous agonists and partial antagonists that is similar to observed increase in Kd (Zhang et al., Mol. Endo., 10, 24-34 (1996)).

References

787I

X:

Confict in Xenopus sequence S or P (P49844)

788K

K to R in marmoset is one of 4 mutations in steroid binding domain proposed to be responsible for 10 fold lower affinity (Brandon et al. 1991).

References

792F

M:

F to A caused 7 fold decreased activity and affinity with TA and increased reductions with other steroids

SM

F to L caused a 5 fold decreased affinity in cell free translated receptor ((Reynolds et al., J. Clin. Endo. Met., 82:465-472 (1997))

793H

M:

H to L has no effect on biological activity (personal communication: Michael Stallcup [USC, Calif.])

M:

H to L has no effect on transcriptional activity (Chen et al., Mol. Endo. 8:422-430[1994]).

References