FRINK Linked Data Fragments server

Matches in SemOpenAlex for { <https://semopenalex.org/work/W2032250517> ?p ?o ?g. }

• W2032250517 endingPage "27956" @default.
• W2032250517 startingPage "27949" @default.
• W2032250517 abstract "In this work, we provide a rationale for the finding that the estrogen receptor (ER) binds to its DNA response element as a homodimer in vivo. Binding of the monomer estrogen receptor DNA binding domain (ER DBD) to a palindromic, consensus estrogen response element (ERE) is increased 5–6-fold when the ER DBD is dimerized either by a monoclonal antibody that recognizes an attached epitope tag or by expressing the ER DBD as a single molecule in which the two monomers are joined by a peptide linker. Most of the increase in binding is due to stabilization of the ER DBD·ERE complex. We observed only an approximately 2.5-fold reduction in binding when a consensus ERE was replaced with widely spaced ERE half-sites, suggesting that the interaction between ER DBDs on the ERE is relatively weak, and that in full-length ER the DBDs can move independently of each other. To test binding to an imperfect palindrome, typical of the imperfect EREs found in almost all natural estrogen receptor responsive genes, we used the pS2 ERE. Even at high concentrations of ER DBD, specific binding of the ER DBD to the imperfect pS2 ERE was undetectable. Both of the dimerized ER DBDs exhibited efficient binding to the imperfect pS2 ERE, with an affinity at least 25-fold greater than monomer ER DBD. These data support the view that steroid receptor dimerization provides an important mechanism facilitating the recognition of naturally occurring, imperfect hormone response elements. In this work, we provide a rationale for the finding that the estrogen receptor (ER) binds to its DNA response element as a homodimer in vivo. Binding of the monomer estrogen receptor DNA binding domain (ER DBD) to a palindromic, consensus estrogen response element (ERE) is increased 5–6-fold when the ER DBD is dimerized either by a monoclonal antibody that recognizes an attached epitope tag or by expressing the ER DBD as a single molecule in which the two monomers are joined by a peptide linker. Most of the increase in binding is due to stabilization of the ER DBD·ERE complex. We observed only an approximately 2.5-fold reduction in binding when a consensus ERE was replaced with widely spaced ERE half-sites, suggesting that the interaction between ER DBDs on the ERE is relatively weak, and that in full-length ER the DBDs can move independently of each other. To test binding to an imperfect palindrome, typical of the imperfect EREs found in almost all natural estrogen receptor responsive genes, we used the pS2 ERE. Even at high concentrations of ER DBD, specific binding of the ER DBD to the imperfect pS2 ERE was undetectable. Both of the dimerized ER DBDs exhibited efficient binding to the imperfect pS2 ERE, with an affinity at least 25-fold greater than monomer ER DBD. These data support the view that steroid receptor dimerization provides an important mechanism facilitating the recognition of naturally occurring, imperfect hormone response elements. The intracellular actions of estrogens are mediated by the estrogen receptor. The estrogen receptor (ER) 1The abbreviations used are: ER, estrogen receptor; ER DBD, estrogen receptor DNA binding domain; AbD-DBD, antibody-dimerized estrogen receptor DNA binding domain; LD-DBD, linker-dimerized estrogen receptor DNA binding domain; PAL3, palindrome with 3 nucleotides between ERE half-sites; EV15, everted ERE with 15 nucleotides between half-sites; DR15, direct repeat of ERE half-sites separated by 15 nucleotides; PCR, polymerase chain reaction; GR, glucocorticoid receptor; ERE, estrogen response element. 1The abbreviations used are: ER, estrogen receptor; ER DBD, estrogen receptor DNA binding domain; AbD-DBD, antibody-dimerized estrogen receptor DNA binding domain; LD-DBD, linker-dimerized estrogen receptor DNA binding domain; PAL3, palindrome with 3 nucleotides between ERE half-sites; EV15, everted ERE with 15 nucleotides between half-sites; DR15, direct repeat of ERE half-sites separated by 15 nucleotides; PCR, polymerase chain reaction; GR, glucocorticoid receptor; ERE, estrogen response element. and other members of the steroid/nuclear receptor superfamily of transcription factors share a common domain structure with discrete regions of the receptors responsible for ligand binding, DNA binding, dimerization, nuclear localization, and transcription activation. Two estrogen receptor domains play a role in dimerization. A strong dimerization interface is located in the ligand binding domain of nuclear receptors (1White R. Fawell S.E. Parker M.G. J. Steroid Biochem. Mol. Biol. 1991; 40: 333-341Crossref PubMed Scopus (16) Google Scholar). A weak dimerization interface is located in the DNA binding domain of steroid receptors (2Schwabe J.W.R. Chapman L. Finch J.T. Rhodes D. Cell. 1993; 75: 567-578Abstract Full Text PDF PubMed Scopus (588) Google Scholar, 3Kumar V. Chambon P. Cell. 1988; 55: 145-156Abstract Full Text PDF PubMed Scopus (956) Google Scholar). However, this interface is insufficient to induce solution dimerization, inasmuch as even at high protein concentrations the isolated recombinant ER DNA binding domain is a monomer in solution (4Nardulli A.M. Lew D. Erijman L. Shapiro D.J. J. Biol. Chem. 1991; 266: 24070-24076Abstract Full Text PDF PubMed Google Scholar, 5Schwabe J.W.R. Neuhaus D. Rhodes D. Nature. 1990; 348: 458-461Crossref PubMed Scopus (348) Google Scholar). When two ER DBD monomers bind to the half-sites of an estrogen response element (ERE), a dimerization interface stabilizes binding of the DBD monomers to the response element (2Schwabe J.W.R. Chapman L. Finch J.T. Rhodes D. Cell. 1993; 75: 567-578Abstract Full Text PDF PubMed Scopus (588) Google Scholar). We refer to this as ERE-dependent DBD dimerization. Although the ER DBD is a monomer in solution, in this work, we did not observe stable occupancy of only one of the two ERE half-sites by a single ER DBD. The ER DBD monomers on the ERE occupy both half-sites and are in the form of ERE-dimerized DBD. The DNA-dependent dimerization interface in the glucocorticoid receptor (GR)-DBD is sufficiently stable that it can alter binding of GR-DBD monomers to a glucocorticoid response element with an incorrect 4-base spacing (6Luisi B.F. Xu W.X. Otwinowski Z. Freedman L.P. Yamamoto K.R. Sigler P.B. Nature. 1991; 352: 497-505Crossref PubMed Scopus (1217) Google Scholar).It is widely, but not universally (7Murdoch F.E. Byrne L.M. Ariazi E.A. Furlow J.D. Meier D.A. Gorski J. Biochemistry. 1995; 34: 9144-9150Crossref PubMed Scopus (41) Google Scholar), accepted that steroid receptors bind to their DNA recognition sequences as dimers. However, ligand binding domain-mediated dimerization is not always required for binding of steroid receptors to their hormone response elements. For example, the ER DBD monomer expressed in Escherichia coli binds to the ERE with a reduced but still significant affinity (3Kumar V. Chambon P. Cell. 1988; 55: 145-156Abstract Full Text PDF PubMed Scopus (956) Google Scholar). It has been widely assumed, but never actually demonstrated, that dimerization of steroid receptors increases their affinity for their DNA response elements.In this work, we address the effect of dimerization on the interaction of a steroid receptor with a hormone response element, and provide a biological rationale for receptor dimerization. Although ER mutants that are impaired in ligand binding domain dimerization have been reported (1White R. Fawell S.E. Parker M.G. J. Steroid Biochem. Mol. Biol. 1991; 40: 333-341Crossref PubMed Scopus (16) Google Scholar), it appears that these mutants still exhibit some ability to dimerize (8Schodin D.J. Zhuang Y. Shapiro D.J. Katzenellenbogen B.S. J. Biol. Chem. 1995; 270: 31163-31171Abstract Full Text Full Text PDF PubMed Scopus (60) Google Scholar). We therefore elected to use the ER DBD, which has been shown clearly to be a monomer in solution (4Nardulli A.M. Lew D. Erijman L. Shapiro D.J. J. Biol. Chem. 1991; 266: 24070-24076Abstract Full Text PDF PubMed Google Scholar, 5Schwabe J.W.R. Neuhaus D. Rhodes D. Nature. 1990; 348: 458-461Crossref PubMed Scopus (348) Google Scholar). We used two very different techniques to artificially dimerize the ER DBD in solution and compared binding to EREs by the dimerized DBD and by the ER DBD monomer. By measuring both equilibrium binding and the association and dissociation of the ER DBDs from a consensus ERE, we show that most of the increase in binding seen with the dimerized ER DBDs is to due to stabilization of the dimerized ER DBD complexes on the ERE. Virtually all EREs identified in ER-regulated genes contain imperfect ERE palindromes. The ER DBD was unable to recognize and bind to an imperfect, non-consensus ERE (from the human PS2 gene), whereas the artificially dimerized ER DBDs bound with high affinity. The striking dimerization-dependent enhancement of binding to an imperfect ERE provides a rationale for dimerization of the estrogen receptor and the other steroid receptors.DISCUSSIONThe DNA recognition sequences of the estrogen receptor and the other steroid hormone receptors are usually palindromic half-sites. In principle, the steroid receptors could interact with their half-sites by binding to them independently as monomers, by binding as monomers and forming interacting dimers on the DNA, or by forming dimers in solution and binding to their recognition sequences as dimeric units. Although several studies have demonstrated that the isolated ER and GR DNA binding domains are monomers in solution and undergo DNA dependent-dimerization after binding to their recognition sequences (4Nardulli A.M. Lew D. Erijman L. Shapiro D.J. J. Biol. Chem. 1991; 266: 24070-24076Abstract Full Text PDF PubMed Google Scholar,18Dahlman-Wright K. Wright A. Gustafsson J. Carlstedt-Duke J. J. Biol. Chem. 1991; 266: 3107-3112Abstract Full Text PDF PubMed Google Scholar), most evidence supports the view that the full-length liganded ER, GR, and progesterone receptor dimerize in solution and bind to their response elements as dimers (reviewed in Ref. 19Glass C. Endocr. Rev. 1994; 15: 391-407PubMed Google Scholar). The recent observation that widely separated ERE half-sites can bind the ER and play a role in transcription activation (14Kato S. Tora L. Yamauchi J. Masushige S. Bellard M. Chambon P. Cell. 1992; 68: 731-742Abstract Full Text PDF PubMed Scopus (268) Google Scholar, 15Kato S. Sasaki H. Suzawa M. Masushige S. Tora L. Chambon P. Gronemeyer H. Mol. Cell. Biol. 1995; 15: 5858-5867Crossref PubMed Scopus (146) Google Scholar) underscores the question of whether dimerization actually confers a significant advantage to the receptor in forming a complex with its hormone response element. In the absence of any quantitative determination of the extent to which receptor dimerization results in enhanced binding to the hormone response element, there was no explanation of the advantage conferred on the receptor by dimerizing.One approach to evaluating the effect of dimerization on interaction of the ER with the ERE would be to compare ERE binding by wild-type ER and by an ER mutant unable to dimerize. Although several ER mutants that show impaired dimerization on translation in a rabbit reticulocyte lysate have been reported (20Fawell S.E. Lees J.A. White R. Parker M.G. Cell. 1990; 60: 953-962Abstract Full Text PDF PubMed Scopus (483) Google Scholar), these mutants likely still retain some ability to dimerize in vivo. For example, one well characterized mutant in this group, L507R, retains significant ability to activate transcription from EREs (8Schodin D.J. Zhuang Y. Shapiro D.J. Katzenellenbogen B.S. J. Biol. Chem. 1995; 270: 31163-31171Abstract Full Text Full Text PDF PubMed Scopus (60) Google Scholar), and is therefore likely to exhibit significant dimerization in vivo. We therefore elected to investigate binding of the ER DNA binding domain to the ERE. The ER DBD has been shown unambiguously to be a monomer in solution (4Nardulli A.M. Lew D. Erijman L. Shapiro D.J. J. Biol. Chem. 1991; 266: 24070-24076Abstract Full Text PDF PubMed Google Scholar,5Schwabe J.W.R. Neuhaus D. Rhodes D. Nature. 1990; 348: 458-461Crossref PubMed Scopus (348) Google Scholar), its interaction with the ERE has been described at high resolution (2Schwabe J.W.R. Chapman L. Finch J.T. Rhodes D. Cell. 1993; 75: 567-578Abstract Full Text PDF PubMed Scopus (588) Google Scholar), and it is available in pure form.To artificially dimerize the ER DBD, we used a peptide linker related to those used in the production of single chain antibodies. Similar linkers have recently been used in a few studies of DNA-binding proteins (21Liang H. Sandberg W.S. Terwilliger T.C. Proc. Natl. Acad. Sci. U. S. A. 1993; 90: 7010-7014Crossref PubMed Scopus (54) Google Scholar, 22Percipalle P. Simoncsits A. Zakhariev S. Guarnaccia C. Sanchez R. Pongor S. EMBO J. 1995; 14: 3200-3205Crossref PubMed Scopus (23) Google Scholar, 23Pomerantz J.L. Sharp P.A. Pabo C.O. Science. 1995; 267: 93-96Crossref PubMed Scopus (121) Google Scholar, 24Robinson C.R. Sauer R.T. Biochemistry. 1996; 35: 109-116Crossref PubMed Scopus (48) Google Scholar). In one such study, the possibility that a peptide linker very similar to the one we used enhanced DNA binding by altering the structure of the protein was explicitly examined (24Robinson C.R. Sauer R.T. Biochemistry. 1996; 35: 109-116Crossref PubMed Scopus (48) Google Scholar). Fluorescence emission, circular dichroism, and DNase I and copper phenanthroline footprinting all indicated that the free arc repressor, and the arc repressor dimerized with a peptide linker had similar structures and interactions with DNA. Because none of these techniques could prove definitively that enhanced binding of the LD-DBD to the ERE was caused by dimerization and not by a linker-mediated structural change in the ER DBD, we also used a second entirely different antibody-based method to dimerize the ER DBD.Although it has been reported previously that antibodies to ER can enhance dimerization (20Fawell S.E. Lees J.A. White R. Parker M.G. Cell. 1990; 60: 953-962Abstract Full Text PDF PubMed Scopus (483) Google Scholar, 25Nardulli A.M. Greene G.L. Shapiro D.J. Mol. Endocrinol. 1993; 7: 331-340Crossref PubMed Scopus (61) Google Scholar), quantitative studies of the interaction of an antibody-dimerized protein with its recognition sequence have not been reported. To dimerize the ER DBD, we used a monoclonal antibody to a short epitope added to the N terminus of the DBD. The short N-terminal flag epitope did not alter DNA binding by the ER DBD as the flag-ER DBD and DBD from which the flag epitope was removed by thrombin digestion showed similar binding to the ERE in gel-shift assays. The position of the epitope recognized by the antibody does not appear to be critical for antibody-mediated dimerization to enhance binding to the ERE. In qualitative studies, we also observed a substantial enhancement of ERE binding using the Xenopus ER DBD and the monoclonal antibody p1A3, which appears to recognize an epitope in the hinge region of the Xenopus ER DBD (data not shown). Inasmuch as this region is poorly conserved, this antibody does not bind to the human ER DBD, necessitating use of the XenopusER DBD for these studies.In contrast to the LD-DBD, which will be a dimer independent of concentration, the antibody will only dimerize the DBD at the appropriate ratio of antibody and flag-DBD antigen. In practice, we found that the DBD bound to the ERE as an antibody-induced dimer across a wide range of antibody:DBD ratios (Fig. 3). One possible explanation for this is that when the antibody has dimerized the DBD, antibody-DBD complex binds rapidly to the consensus ERE (Fig. 5 B), and once bound the ERE may be stabilizing the antibody complex with two bound DBDs. Support for this idea comes from the data for the AbD-DBD bound to the non-consensus pS2 ERE. Qualitative assessment of the amount of probe up-shifted at the three antibody concentrations indicates more variation than was seen for the consensus ERE.Most available data indicate that the liganded full-length ER is a dimer in solution. However, the state of the DBDs in the solution dimer of ER is unknown. It is not clear whether the DBDs in the solution dimer of ER are linked to the rest of the protein through a conformationally flexible region (as has been proposed for nuclear receptors; Ref. 26Luisi B. Freedman L. Nature. 1995; 375: 359-360Crossref PubMed Scopus (9) Google Scholar) and are therefore free to move independently, or are associated through the dimerization interface seen in the x-ray structure of two ER DBDs bound to the ERE. The observation that an ERE with a direct repeat of the ERE half-sites with 15 nucleotides between the half-sites bound the ER and activated transcription approximately one-third as well as the consensus ERE palindrome (15Kato S. Sasaki H. Suzawa M. Masushige S. Tora L. Chambon P. Gronemeyer H. Mol. Cell. Biol. 1995; 15: 5858-5867Crossref PubMed Scopus (146) Google Scholar) raised the possibility that the DBDs in the intact protein are not in a dimer interface but are free to rotate. To evaluate this question, we determined the effect of dimerization on binding of ER DBD to the consensus ERE palindrome with its 3 nucleotide spacer (PAL3) and to direct repeats (DR15) and everted repeats (EV15) with a 15-nucleotide spacer. Binding of LD-DBD to DR15 was inefficient, presumably because the short length of the linker required the introduction of an energetically unfavorable bend or kink in the DNA. In contrast, LD-DBD bound to EV15 with only about a 2.5-fold lower affinity than to PAL3 (Table I). Because the two DBD monomers are separated by an extra turn of the DNA helix and are in opposite orientation when bound to EV15, they are unlikely to form the DNA-dependent dimerization interface. The 2.5-fold difference in binding of LD-DBD to PAL3 and EV15 corresponds to a DBD dimerization energy of about 0.5 kcal/mol, which is approximately 4% of the 12 kcal/mol total energy of interaction of the full-length ER with the consensus ERE (which has been determined to have a dissociation constant of 2.0 ± 0.3 nm; Ref. 27Carlsson B. Haggblad J. Anal. Biochem. 1995; 232: 172-179Crossref PubMed Scopus (18) Google Scholar). This relatively small contribution of DBD dimerization to the total binding energy implies that the DBDs in dimerized ER may be free to rotate and bind to functional EREs, which are not palindromes and do not have the familiar three-base pair spacing. Furthermore, our observation raises the possibility that widely spaced everted ERE half-sites may similarly function in vivo, although none have yet been reported.Table IDissociation constants for DBD construct and ERE probe combinationsERE probeDBD constructDissociation constant1-aThe K D values were calculated from the data presented in Figs. 2, 3, 4, 6, and 7.nmConsensus EREDBD160LD-DBD38AbD-DBD29EV15LD-DBD∼100pS2 nonconsensus EREDBDUndetectableLD-DBD110AbD-DBD∼2001-a The K D values were calculated from the data presented in Figs. 2, 3, 4, 6, and 7. Open table in a new tab Our kinetics studies examine two possible mechanisms by which the ER may bind to the ERE. Binding of the ER to the ERE may occur by a sequential mechanism in which one DBD binds followed by binding of the second DBD. Because binding of a single DBD to an ERE half-site is strongly disfavored (28Green S. Chambon P. Parker M.G. Nuclear Hormone Receptors. Academic Press, New York1991: 15-38Google Scholar), the second DBD must bind to one ERE half-site before the first DBD has dissociated from the other half-site. Alternatively, the two DBDs may transiently dimerize in solution and then rapidly bind to the ERE as a unit. For either of these mechanisms, we would expect an increase in the on-rate in response to externally induced dimerization of the DBD, such as that brought about by sequences in the ligand binding domain. In the case of the sequential mechanism, this would be due to the dimerization-induced high local concentration of the second DBD once the first DBD was bound. In the transient dimerization model, only a small proportion of the DBD is a dimer at any one time and dimerizing all of the DBD via sequences in the ligand binding domain would increase the proportion of DBD in the ER-binding dimer form.We find that for the AbD-DBD binding is extremely rapid, even at 4 °C. However, the LD-DBD actually has a slower on-rate than the monomer DBD. This may be due to the linker interfering with binding or to the presence of the linker (which is attached near the outsides of the protein complex when bound to a consensus ERE) favoring a solution form of the linked DBD monomers in which the two DBDs are as far apart as possible and therefore have an orientation opposite to the orientation needed for binding.It is the rate of dissociation from the ERE (the off-rate) that is most strongly affected by dimerizing the DBD. If the ER DBD monomers bind and dissociate sequentially from the DBD, the DBD·ERE complex is stabilized by the relatively weak dimerization interface between the two monomers on the ERE, and the interaction of each monomer with a half-site. Because binding of the ER or the ER DBD to a single half-site is not favored (28Green S. Chambon P. Parker M.G. Nuclear Hormone Receptors. Academic Press, New York1991: 15-38Google Scholar), dissociation of one DBD monomer results in an unstable complex of one DBD and an ERE-half-site. In contrast, when the DBDs are dimerized, one DBD would maintain transient contact with an ERE half-site after the other monomer had dissociated. Dimerizing the DBD would greatly increase the local ER DBD concentration and the probability that the monomer that had dissociated would rebind to the free ERE half-site. This view is consistent with the data showing that dimerization produces a dramatic increase in binding to an imperfect ERE. When one half-site contains an imperfect ERE, as is found in the pS2 ERE, the energetic contribution due to binding of the single ERE monomer is insufficient for forming a stable complex and no binding is observed. The inability of a single ER DBD monomer to bind to a half-site under these conditions is illustrated by the smear (presumably due to a rapid dissociation of half-site complexes) observed when the pS2 ERE is incubated with extremely high levels of ER DBD. The absence of a complex of ER DBD on both half-sites of the pS2 ERE is consistent with our view that the dimerization interface between the ER DBDs bound to the ERE is relatively weak and cannot stabilize binding of the DBD to the imperfect half-site.Dimerizing the DBD produced a dramatic increase in binding to the non-consensus pS2 ERE, which differs from the ERE consensus sequence by only one nucleotide in one half-site. It was not possible to accurately quantitate the effect of dimerization on binding to the pS2 ERE, inasmuch as even at a concentration of 10,000 nm, the DBD monomer did not produce a discrete upshifted band in the gel mobility shift assay with the pS2 probe. At the highest concentrations used, there was some smearing near the probe alone band. Even if this smearing is considered to represent binding of the ER DBD to the ERE, the affinity of the dimerized DBDs is at least 25-fold higher than the affinity of the DBD monomer. This is a substantially larger effect than the 5–6-fold difference in binding seen with the consensus ERE palindrome. This suggests that dimerization stabilizes the interaction of the DBD with the imperfect ERE half-site. The imperfect pS2 ERE binds ER with only a 2–5-fold lower affinity than the consensus ERE (3Kumar V. Chambon P. Cell. 1988; 55: 145-156Abstract Full Text PDF PubMed Scopus (956) Google Scholar, 29Nardulli A.M. Romine L.E. Carpo C. Greene G.L. Rainish B. Mol. Endocrinol. 1996; 10: 694-704PubMed Google Scholar). The striking effect of dimerization on binding to the pS2 ERE suggests that for both the pS2 ERE and for the many other EREs that exhibit larger deviations from the consensus sequence, ER dimerization is a prerequisite for efficient ER binding.In this work, we used two independent methods to quantitate the effect of dimerization on binding of the ER DBD to the ERE. Surprisingly, the major effect of dimerization was to stabilize the ER DBD·ERE complex rather than to increase the association rate. Our data support the view that, in the intact ER dimer, the DBDs can behave independently of each other. Most important, dimerization was required for binding to an imperfect naturally occurring ERE. These data support a model in which the ER must be dimerized to occupy both ERE half-sites in the imperfect EREs found in almost all ER-regulated genes. The intracellular actions of estrogens are mediated by the estrogen receptor. The estrogen receptor (ER) 1The abbreviations used are: ER, estrogen receptor; ER DBD, estrogen receptor DNA binding domain; AbD-DBD, antibody-dimerized estrogen receptor DNA binding domain; LD-DBD, linker-dimerized estrogen receptor DNA binding domain; PAL3, palindrome with 3 nucleotides between ERE half-sites; EV15, everted ERE with 15 nucleotides between half-sites; DR15, direct repeat of ERE half-sites separated by 15 nucleotides; PCR, polymerase chain reaction; GR, glucocorticoid receptor; ERE, estrogen response element. 1The abbreviations used are: ER, estrogen receptor; ER DBD, estrogen receptor DNA binding domain; AbD-DBD, antibody-dimerized estrogen receptor DNA binding domain; LD-DBD, linker-dimerized estrogen receptor DNA binding domain; PAL3, palindrome with 3 nucleotides between ERE half-sites; EV15, everted ERE with 15 nucleotides between half-sites; DR15, direct repeat of ERE half-sites separated by 15 nucleotides; PCR, polymerase chain reaction; GR, glucocorticoid receptor; ERE, estrogen response element. and other members of the steroid/nuclear receptor superfamily of transcription factors share a common domain structure with discrete regions of the receptors responsible for ligand binding, DNA binding, dimerization, nuclear localization, and transcription activation. Two estrogen receptor domains play a role in dimerization. A strong dimerization interface is located in the ligand binding domain of nuclear receptors (1White R. Fawell S.E. Parker M.G. J. Steroid Biochem. Mol. Biol. 1991; 40: 333-341Crossref PubMed Scopus (16) Google Scholar). A weak dimerization interface is located in the DNA binding domain of steroid receptors (2Schwabe J.W.R. Chapman L. Finch J.T. Rhodes D. Cell. 1993; 75: 567-578Abstract Full Text PDF PubMed Scopus (588) Google Scholar, 3Kumar V. Chambon P. Cell. 1988; 55: 145-156Abstract Full Text PDF PubMed Scopus (956) Google Scholar). However, this interface is insufficient to induce solution dimerization, inasmuch as even at high protein concentrations the isolated recombinant ER DNA binding domain is a monomer in solution (4Nardulli A.M. Lew D. Erijman L. Shapiro D.J. J. Biol. Chem. 1991; 266: 24070-24076Abstract Full Text PDF PubMed Google Scholar, 5Schwabe J.W.R. Neuhaus D. Rhodes D. Nature. 1990; 348: 458-461Crossref PubMed Scopus (348) Google Scholar). When two ER DBD monomers bind to the half-sites of an estrogen response element (ERE), a dimerization interface stabilizes binding of the DBD monomers to the response element (2Schwabe J.W.R. Chapman L. Finch J.T. Rhodes D. Cell. 1993; 75: 567-578Abstract Full Text PDF PubMed Scopus (588) Google Scholar). We refer to this as ERE-dependent DBD dimerization. Although the ER DBD is a monomer in solution, in this work, we did not observe stable occupancy of only one of the two ERE half-sites by a single ER DBD. The ER DBD monomers on the ERE occupy both half-sites and are in the form of ERE-dimerized DBD. The DNA-dependent dimerization interface in the glucocorticoid receptor (GR)-DBD is sufficiently stable that it can alter binding of GR-DBD monomers to a glucocorticoid response element with an incorrect 4-base spacing (6Luisi B.F. Xu W.X. Otwinowski Z. Freedman L.P. Yamamoto K.R. Sigler P.B. Nature. 1991; 352: 497-505Crossref PubMed Scopus (1217) Google Scholar). It is widely, but not universally (7Murdoch F.E. Byrne L.M. Ariazi E.A. Furlow J.D. Meier D.A. Gorski J. Biochemistry. 1995; 34: 9144-9150Crossref PubMed Scopus (41) Google Scholar), accepted that steroid receptors bind to their DNA recognition sequences as dimers. However, ligand binding domain-mediated dimerization is not always required for binding of steroid receptors to their hormone response elements. For example, the ER DBD monomer expressed in Escherichia coli binds to the ERE with a reduced but still significant affinity (3Kumar V. Chambon P. Cell. 1988; 55: 145-156Abstract Full Text PDF PubMed Scopus (956) Google Scholar). It has been widely assumed, but never actually demonstrated, that dimerization of steroid receptors increases their affinity for their DNA response elements. In this work, we address the effect of dimerization on the interaction of a steroid receptor with a hormone response element, and provide a biological rationale for receptor dimerization. Although ER mutants that are impaired in ligand binding domain dimerization have been reported (1White R. Fawell S.E. Parker M.G. J. Steroid Biochem. Mol. Biol. 1991; 40: 333-341Crossref PubMed Scopus (16) Google Scholar), it appears that these mutants still exhibit some ability to dimerize (8Schodin D.J. Zhuang Y. Shapiro D.J. Katzenellenbogen B.S. J. Biol. Chem. 1995; 270: 31163-31171Abstract Full Text Full Text PDF PubMed Scopus (60) Google Scholar). We therefore elected to use the ER DBD, which has been shown clearly to be a monomer in solution (4Nardulli A.M. Lew D. Erijman L. Shapiro D.J. J. Biol. Chem. 1991; 266: 24070-24076Abstract Full Text PDF PubMed Google Scholar, 5Schwabe J.W.R. Neuhaus D. Rhodes D. Nature. 1990; 348: 458-461Crossref PubMed Scopus (34" @default.
• W2032250517 created "2016-06-24" @default.
• W2032250517 creator A5030190949 @default.
• W2032250517 creator A5061599949 @default.
• W2032250517 date "1997-10-01" @default.
• W2032250517 modified "2023-10-13" @default.
• W2032250517 title "Dimerizing the Estrogen Receptor DNA Binding Domain Enhances Binding to Estrogen Response Elements" @default.
• W2032250517 cites W1489022720 @default.
• W2032250517 cites W1534882737 @default.
• W2032250517 cites W1585759343 @default.
• W2032250517 cites W1968699661 @default.
• W2032250517 cites W1969820155 @default.
• W2032250517 cites W1978313171 @default.
• W2032250517 cites W1983902996 @default.
• W2032250517 cites W1997007589 @default.
• W2032250517 cites W2001951212 @default.
• W2032250517 cites W2007705662 @default.
• W2032250517 cites W2013700099 @default.
• W2032250517 cites W2014107736 @default.
• W2032250517 cites W2018579779 @default.
• W2032250517 cites W2022704908 @default.
• W2032250517 cites W2028605407 @default.
• W2032250517 cites W2041781038 @default.
• W2032250517 cites W2055650366 @default.
• W2032250517 cites W2066441904 @default.
• W2032250517 cites W2100837269 @default.
• W2032250517 cites W2132185105 @default.
• W2032250517 cites W2143678523 @default.
• W2032250517 cites W2155594578 @default.
• W2032250517 cites W2186971407 @default.
• W2032250517 doi "https://doi.org/10.1074/jbc.272.44.27949" @default.
• W2032250517 hasPubMedId "https://pubmed.ncbi.nlm.nih.gov/9346945" @default.
• W2032250517 hasPublicationYear "1997" @default.
• W2032250517 type Work @default.
• W2032250517 sameAs 2032250517 @default.
• W2032250517 citedByCount "44" @default.
• W2032250517 countsByYear W20322505172012 @default.
• W2032250517 countsByYear W20322505172013 @default.
• W2032250517 countsByYear W20322505172017 @default.
• W2032250517 countsByYear W20322505172018 @default.
• W2032250517 countsByYear W20322505172019 @default.
• W2032250517 countsByYear W20322505172020 @default.
• W2032250517 countsByYear W20322505172022 @default.
• W2032250517 countsByYear W20322505172023 @default.
• W2032250517 crossrefType "journal-article" @default.
• W2032250517 hasAuthorship W2032250517A5030190949 @default.
• W2032250517 hasAuthorship W2032250517A5061599949 @default.
• W2032250517 hasBestOaLocation W20322505171 @default.
• W2032250517 hasConcept C104317684 @default.
• W2032250517 hasConcept C107824862 @default.
• W2032250517 hasConcept C121608353 @default.
• W2032250517 hasConcept C134018914 @default.
• W2032250517 hasConcept C185592680 @default.
• W2032250517 hasConcept C25166345 @default.
• W2032250517 hasConcept C2777164284 @default.
• W2032250517 hasConcept C33987129 @default.
• W2032250517 hasConcept C51445715 @default.
• W2032250517 hasConcept C530470458 @default.
• W2032250517 hasConcept C54355233 @default.
• W2032250517 hasConcept C552990157 @default.
• W2032250517 hasConcept C55493867 @default.
• W2032250517 hasConcept C84606932 @default.
• W2032250517 hasConcept C86339819 @default.
• W2032250517 hasConcept C86803240 @default.
• W2032250517 hasConcept C95444343 @default.
• W2032250517 hasConcept C96307122 @default.
• W2032250517 hasConceptScore W2032250517C104317684 @default.
• W2032250517 hasConceptScore W2032250517C107824862 @default.
• W2032250517 hasConceptScore W2032250517C121608353 @default.
• W2032250517 hasConceptScore W2032250517C134018914 @default.
• W2032250517 hasConceptScore W2032250517C185592680 @default.
• W2032250517 hasConceptScore W2032250517C25166345 @default.
• W2032250517 hasConceptScore W2032250517C2777164284 @default.
• W2032250517 hasConceptScore W2032250517C33987129 @default.
• W2032250517 hasConceptScore W2032250517C51445715 @default.
• W2032250517 hasConceptScore W2032250517C530470458 @default.
• W2032250517 hasConceptScore W2032250517C54355233 @default.
• W2032250517 hasConceptScore W2032250517C552990157 @default.
• W2032250517 hasConceptScore W2032250517C55493867 @default.
• W2032250517 hasConceptScore W2032250517C84606932 @default.
• W2032250517 hasConceptScore W2032250517C86339819 @default.
• W2032250517 hasConceptScore W2032250517C86803240 @default.
• W2032250517 hasConceptScore W2032250517C95444343 @default.
• W2032250517 hasConceptScore W2032250517C96307122 @default.
• W2032250517 hasIssue "44" @default.
• W2032250517 hasLocation W20322505171 @default.
• W2032250517 hasOpenAccess W2032250517 @default.
• W2032250517 hasPrimaryLocation W20322505171 @default.
• W2032250517 hasRelatedWork W1521366099 @default.
• W2032250517 hasRelatedWork W1971423810 @default.
• W2032250517 hasRelatedWork W2009445334 @default.
• W2032250517 hasRelatedWork W2024832428 @default.
• W2032250517 hasRelatedWork W2032095915 @default.
• W2032250517 hasRelatedWork W2032250517 @default.
• W2032250517 hasRelatedWork W2092903481 @default.
• W2032250517 hasRelatedWork W2951509230 @default.
• W2032250517 hasRelatedWork W4253637622 @default.
• W2032250517 hasRelatedWork W171128861 @default.

• next