PMID- 10571988
OWN - NLM
STAT- MEDLINE
DCOM- 19991217
LR  - 20190613
IS  - 0006-2960 (Print)
IS  - 0006-2960 (Linking)
VI  - 38
IP  - 43
DP  - 1999 Oct 26
TI  - Function of estrogen receptor tyrosine 537 in hormone binding, DNA binding, and
      transactivation.
PG  - 14146-56
AB  - The human estrogen receptor (hER) is a ligand-activated transcription factor
      which, like many other members of the nuclear receptor protein family, exhibits a
      dimerization-dependent transcriptional activation. Several previous reports have 
      provided evidence of the phosphorylation of the hER at tyrosine 537 (Y537).
      However, the exact function of a putative phosphorylation at this site remains
      controversial. Using a yeast transactivation assay, and in vitro biochemical
      approaches, we show that phosphorylation of tyrosine 537 is not required for the 
      hER to bind hormone, or to activate transcription. An hER tyrosine 537 to
      phenylalanine (Y537F) mutant retains 70-75% of the transactivation potential of
      wild type hER in a yeast reporter system. Furthermore, the mutated receptor
      exhibits wild type hormone and DNA binding affinities. However, this mutation
      results in a decrease in receptor stability as measured by a decrease in the
      extent of hormone binding over time. The most striking difference between the
      wild type and Y537F hER is in the estradiol binding kinetics. Whereas the
      off-rate for estradiol exhibits a two-state binding mechanism, the Y537F mutant
      hER exhibits a monophasic estradiol off-rate. On the basis of these data and
      other reports describing the structure and activity of Y537 mutations, as well as
      knowledge of the three-dimensional structure of the hER ligand binding domain, we
      propose an alternate model wherein Y537F mutation favors an "open" pocket
      conformation, affecting the estrogen binding kinetics and stability of the
      hormone-bound, transcriptionally active "closed" pocket conformation. Although
      its phosphorylation is not essential for function of the hER, Y537 is
      nevertheless a critical residue intricately involved with the conformational
      changes of the hER and its ability to activate transcription.
FAU - Yudt, M R
AU  - Yudt MR
AD  - Department of Biochemistry, University of Rochester School of Medicine and
      Dentistry, New York 14642, USA.
FAU - Vorojeikina, D
AU  - Vorojeikina D
FAU - Zhong, L
AU  - Zhong L
FAU - Skafar, D F
AU  - Skafar DF
FAU - Sasson, S
AU  - Sasson S
FAU - Gasiewicz, T A
AU  - Gasiewicz TA
FAU - Notides, A C
AU  - Notides AC
LA  - eng
GR  - ES01247/ES/NIEHS NIH HHS/United States
GR  - HD06707/HD/NICHD NIH HHS/United States
PT  - Comparative Study
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PT  - Research Support, U.S. Gov't, Non-P.H.S.
PT  - Research Support, U.S. Gov't, P.H.S.
PL  - United States
TA  - Biochemistry
JT  - Biochemistry
JID - 0370623
RN  - 0 (DNA-Binding Proteins)
RN  - 0 (Receptors, Estrogen)
RN  - 0 (Recombinant Proteins)
RN  - 0 (Repressor Proteins)
RN  - 42HK56048U (Tyrosine)
RN  - 4TI98Z838E (Estradiol)
SB  - IM
MH  - Amino Acid Sequence
MH  - Animals
MH  - DNA-Binding Proteins/agonists/antagonists & inhibitors/*metabolism/physiology
MH  - Estradiol/genetics/metabolism
MH  - Humans
MH  - Molecular Sequence Data
MH  - Mutagenesis, Site-Directed
MH  - Protein Binding/genetics
MH  - Receptors, Estrogen/agonists/antagonists & inhibitors/genetics/*metabolism
MH  - Recombinant Proteins/agonists/antagonists & inhibitors/genetics/metabolism
MH  - Repressor Proteins/genetics/metabolism
MH  - Response Elements
MH  - *Transcriptional Activation
MH  - Tyrosine/genetics/metabolism/*physiology
EDAT- 1999/11/26 00:00
MHDA- 1999/11/26 00:01
CRDT- 1999/11/26 00:00
PHST- 1999/11/26 00:00 [pubmed]
PHST- 1999/11/26 00:01 [medline]
PHST- 1999/11/26 00:00 [entrez]
AID - bi9911132 [pii]
AID - 10.1021/bi9911132 [doi]
PST - ppublish
SO  - Biochemistry. 1999 Oct 26;38(43):14146-56. doi: 10.1021/bi9911132.