PMID- 8663386
OWN - NLM
STAT- MEDLINE
DCOM- 19960903
LR  - 20210210
IS  - 0021-9258 (Print)
IS  - 0021-9258 (Linking)
VI  - 271
IP  - 30
DP  - 1996 Jul 26
TI  - Identification of amino acids critical for the DNA binding and dimerization 
      properties of the human retinoic acid receptor alpha. Importance of lysine 360, 
      lysine 365, and valine 361.
PG  - 17996-8006
AB  - Retinoic acid receptors (RARs) and retinoid X receptors (RXRs) activate target 
      genes by binding to retinoic acid response elements (RAREs) as heterodimeric, 
      asymmetrical complexes, and display a high degree of cooperativity in binding to 
      RAREs. We have examined here the effect of lysine, cysteine, arginine, histidine, 
      and tyrosine side chain chemical modification on the DNA binding, homo- and 
      heterodimerization properties of the full-length human retinoic acid receptor 
      alpha (hRARalpha). Lysines are the only residues to be engaged in the 
      dimerization with human retinoid X receptor alpha (hRXRalpha) in the absence of 
      DNA, whereas histidines are selectively involved in the homodimerization of 
      hRARalpha in the presence of a RARE. Arginine modification affected the DNA 
      binding activity of each type of dimer, whereas cysteines and tyrosines were 
      primarily involved in the homo- or heterodimerization process in the presence of 
      the same RARE. Modified lysines, interfering with the dimerization with 
      hRXRalpha, were identified by receptor labeling and peptide mapping. They are 
      located in the hormone binding domain eighth heptad repeat, at positions 360 and 
      365. In keeping with these results, mutation of Lys360, Val361, and Lys365 
      diminished strongly the DNA binding activity of hRARalpha as a homodimer or a 
      heterodimer. Our results thus provide direct evidence for the differential 
      involvement of basic, polar, or aromatic amino acids in the DNA binding, 
      homodimerization, and heterodimerization properties of hRARalpha. Furthermore, 
      they demonstrate the use of distinct dimerization interfaces and identify the 
      type of amino acids involved in these protein-protein interactions.
FAU - Rachez, C
AU  - Rachez C
AD  - CJF INSERM 92-03, Laboratoire de Biochimie Structurale, Faculte de Medecine de 
      Lille 1, France.
FAU - Sautiere, P
AU  - Sautiere P
FAU - Formstecher, P
AU  - Formstecher P
FAU - Lefebvre, P
AU  - Lefebvre P
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PL  - United States
TA  - J Biol Chem
JT  - The Journal of biological chemistry
JID - 2985121R
RN  - 0 (Amino Acids)
RN  - 0 (DNA-Binding Proteins)
RN  - 0 (Receptors, Retinoic Acid)
RN  - 0 (Recombinant Proteins)
RN  - 0 (Sulfhydryl Compounds)
RN  - 5688UTC01R (Tretinoin)
SB  - IM
MH  - Amino Acid Sequence
MH  - Amino Acids/chemistry
MH  - DNA-Binding Proteins/chemistry/genetics/*metabolism
MH  - Humans
MH  - Models, Molecular
MH  - Molecular Sequence Data
MH  - Protein Binding
MH  - Protein Conformation
MH  - Receptors, Retinoic Acid/chemistry/genetics/*metabolism
MH  - Recombinant Proteins/chemistry/metabolism
MH  - Structure-Activity Relationship
MH  - Sulfhydryl Compounds/metabolism
MH  - Tretinoin/metabolism
EDAT- 1996/07/26 00:00
MHDA- 1996/07/26 00:01
CRDT- 1996/07/26 00:00
PHST- 1996/07/26 00:00 [pubmed]
PHST- 1996/07/26 00:01 [medline]
PHST- 1996/07/26 00:00 [entrez]
AID - S0021-9258(19)87014-X [pii]
AID - 10.1074/jbc.271.30.17996 [doi]
PST - ppublish
SO  - J Biol Chem. 1996 Jul 26;271(30):17996-8006. doi: 10.1074/jbc.271.30.17996.