Selection of peptides with affinity for the N-terminal domain of GATA-1: Identification of a potential interacting protein.
As most transcription factors, GATA-1 activities are mediated by interactions with multiple proteins. Those identified so far associate with the zinc-finger domain and/or surrounding sequences. In contrast, no proteins interacting with the N-terminal domain have been identified although several evidences suggest its involvement in the control of hematopoiesis. In an attempt to identify proteins that interact with the N-terminal transactivation domain of GATA-1, a random phage peptide library was screened with recombinant GATA-1 protein and the sequence of a selected peptide was used for database protein sequence retrieval. We selected a set of peptides sharing the core sequence phi-B((2-3))-nu((2-4)) (where phi, B, and nu represent hydrophobic, basic, and neutral residues, respectively). Using the sequence of the most represented peptide (pep5) as query, we retrieved the HIV accessory protein Nef. We show that Nef binds GATA-1 and GATA-3 in vitro in virtue of its sequence homology with pep5.
Secco
Paola
P
Department of Medical Sciences and Interdisciplinary Research Center on Autoimmune Diseases, University of Eastern Piedmont A.Avogadro, via Solaroli 17, 28100 Novara, Italy.
Cotella
Diego
D
Santoro
Claudio
C
D000595:Amino Acid Sequence; D000818:Animals; D001665:Binding Sites; D001667:Binding, Competitive; D004268:DNA-Binding Proteins / Q000737:chemistry* / Q000378:metabolism*; D030561:Databases, Nucleic Acid; D050981:Erythroid-Specific DNA-Binding Factors; D050982:GATA1 Transcription Factor; D015693:Gene Products, nef / Q000737:chemistry / Q000378:metabolism*; D051379:Mice; D008969:Molecular Sequence Data; D010455:Peptides / Q000737:chemistry / Q000378:metabolism; D017434:Protein Structure, Tertiary; D011506:Proteins; D014157:Transcription Factors / Q000737:chemistry* / Q000378:metabolism*
D004268:DNA-Binding Proteins; D050981:Erythroid-Specific DNA-Binding Factors; D050982:GATA1 Transcription Factor; C494689:Gata1 protein, mouse; D015693:Gene Products, nef; D010455:Peptides; D011506:Proteins; D014157:Transcription Factors