title: The use of phage display technique for the isolation of androgen receptor interacting peptides with (F/W)XXL(F/W) and FXXLY new signature motifs.
abstract: Early studies suggested that the signature motif, LXXLL, within steroid hormone receptor p160 coregulators may play important roles for the mediation of receptor-coregulator interaction. Interestingly, several androgen receptor (AR) coregulators, such as ARA70 and ARA55, may not use such a unique motif to mediate their coregulator activity. Here we apply the phage display technique to identify some new signature motifs, (F/W)XXL(F/W) and FXXLY (where F is phenylalanine, W is tryptophan, L is leucine, Y is tyrosine, and X is any amino acid) that can influence the interaction between AR and AR coregulators. Sequence analyses found that several AR coregulators, such as ARA70, ARA55, ARA54, and FHL2, contain FXXL(F/Y) motifs. Both glutathione S-transferase pull-down assays and transient transfection reporter assays demonstrate that these AR coregulators may use the FXXL(F/Y) motif to interact with AR and exert their AR coregulator activity. Exchanging the amino acid of Phe, Trp, or Tyr in this newly identified signature motif cluster may influence these peptides to interact with AR. The motif-containing peptides, as well as ARA70 or ARA54, may require selective flanking sequences for the better interaction with AR. In addition to influencing the AR transactivation, these motifs in AR-interacting peptides/proteins were also able to influence the AR N-/C-terminal interaction. Together, our data suggest that AR interacting peptides and/or AR coregulators may utilize the (F/W)XXL(F/W) and FXXLY motifs to mediate their interaction with AR and exert their influences on the AR transactivation.
authors.affiliation: Department of Pathology, University of Rochester Medical Center, Rochester, New York 14642, USA.
authors.forename: Cheng-Lung
authors.initials: CL
authors.lastname: Hsu
authors.forename: Yuh-Ling
authors.initials: YL
authors.lastname: Chen
authors.forename: Shuyuan
authors.initials: S
authors.lastname: Yeh
authors.forename: Huei-Ju
authors.initials: HJ
authors.lastname: Ting
authors.forename: Yueh-Chiang
authors.initials: YC
authors.lastname: Hu
authors.forename: Hank
authors.initials: H
authors.lastname: Lin
authors.forename: Xin
authors.initials: X
authors.lastname: Wang
authors.forename: Chawnshang
authors.initials: C
authors.lastname: Chang
mesh_terms: D020816:Amino Acid Motifs; D000595:Amino Acid Sequence; D001665:Binding Sites; D002352:Carrier Proteins / Q000737:chemistry / Q000378:metabolism; D018398:Homeodomain Proteins / Q000737:chemistry / Q000378:metabolism; D006801:Humans; D047908:Intracellular Signaling Peptides and Proteins; D060588:LIM Domain Proteins; D060850:LIM-Homeodomain Proteins; D009124:Muscle Proteins; D056918:Nuclear Receptor Coactivators; D015513:Oncogene Proteins; D010446:Peptide Fragments / Q000737:chemistry* / Q000378:metabolism; D019151:Peptide Library; D011485:Protein Binding; D011944:Receptors, Androgen / Q000378:metabolism*; D015534:Trans-Activators / Q000737:chemistry / Q000378:metabolism; D014157:Transcription Factors / Q000737:chemistry / Q000378:metabolism
chemical_list: D002352:Carrier Proteins; C105987:FHL2 protein, human; D018398:Homeodomain Proteins; D047908:Intracellular Signaling Peptides and Proteins; D060588:LIM Domain Proteins; D060850:LIM-Homeodomain Proteins; D009124:Muscle Proteins; C111382:NCOA4 protein, human; D056918:Nuclear Receptor Coactivators; D015513:Oncogene Proteins; D010446:Peptide Fragments; D019151:Peptide Library; C118499:RNF14 protein, human; D011944:Receptors, Androgen; C490351:TGFB1I1 protein, human; D015534:Trans-Activators; D014157:Transcription Factors