Imaging analysis of subcellular correlation of androgen receptor and estrogen receptor alpha in single living cells using green fluorescent protein color variants.
Androgen and estrogen act not only in a sex-specific manner but also interactively and synergistically. In the present study, to examine the possible interaction between androgen receptor (AR) and estrogen receptor-alpha (ERalpha), we investigated the subcellular dynamics of AR and ERalpha fused with green fluorescent protein color variants in single living cells using time-lapse microscopy and the technique of fluorescence recovery after photobleaching. AR and ERalpha showed punctate colocalization in the nucleus with estrogen, but not androgen. N-terminal AR deletion mutant did not form a nuclear punctate pattern with either androgen or estrogen. In the presence of AR, but not ERalpha, N-terminal AR deletion mutant formed a punctate nuclear pattern with androgen. AR had different mobility depending on the ligand and the presence of ERalpha. On the other hand, AR had little effect on the stability of ERalpha. ERalpha mutant that does not bind coactivators did not alter the mobility of AR. Taken together, using an imaging technique, we clarified that possible homo/hetero dimerization between AR and ERalpha could be attributed to androgen-estrogen interaction in living cells.
Ochiai
Ikuo
I
Department of Anatomy and Neurobiology, Kyoto Prefectural University of Medicine, Kawaramachi Hirokoji, Kamigyo-ku, Kyoto 602-8566, Japan.
Matsuda
Ken-Ichi
K
Nishi
Mayumi
M
Ozawa
Hitoshi
H
Kawata
Mitsuhiro
M
D000818:Animals; D001426:Bacterial Proteins / Q000032:analysis; D002460:Cell Line; D002467:Cell Nucleus / Q000648:ultrastructure; D002522:Chlorocebus aethiops; D003001:Cloning, Molecular; D047628:Estrogen Receptor alpha; D049452:Green Fluorescent Proteins; D015151:Immunoblotting; D008164:Luminescent Proteins / Q000032:analysis*; D018613:Microscopy, Confocal; D008856:Microscopy, Fluorescence; D016297:Mutagenesis, Site-Directed; D011944:Receptors, Androgen / Q000032:analysis* / Q000235:genetics; D011960:Receptors, Estrogen / Q000032:analysis* / Q000235:genetics; D011993:Recombinant Fusion Proteins / Q000032:analysis; D015183:Restriction Mapping; D014158:Transcription, Genetic
D001426:Bacterial Proteins; D047628:Estrogen Receptor alpha; D008164:Luminescent Proteins; D011944:Receptors, Androgen; D011960:Receptors, Estrogen; D011993:Recombinant Fusion Proteins; C054562:yellow fluorescent protein, Bacteria; D049452:Green Fluorescent Proteins