Molecular and genetic characterization of GABP beta.
This report outlines three observations relating to GABP beta, a polypeptide constituent of the heterotetrameric transcription factor GABP. Evidence is presented showing that the mouse genome encodes two highly related GABP beta polypeptides, designated GABP beta 1-1 and GABP beta 2-1. Genomic and cDNA copies of the newly defined Gabpb2 gene were cloned and characterized, providing the conceptually translated amino acid sequence of GABP beta 2-1. The genes encoding these two proteins, as well as GABP alpha, were mapped to three unlinked chromosomal loci. Although physically unlinked, the patterns of expression of the three genes were strikingly concordant. Finally, the molecular basis of GABP beta dimerization was resolved. Carboxy-terminal regions of the two GABP beta polypeptides, which mediate dimerization, bear highly related primary amino acid sequences. Both sequences are free of alpha-helix destabilizing residues and, when displayed on idealized alpha-helical projections, reveal marked amphipathy. Two observations indicate that these regions adopt an alpha-helical conformation and intertwine as coiled-coils. First, the dimer-forming region of GABP beta 2-1 can functionally replace the leucine zipper of a bZIP transcription factor. Second, a synthetic peptide corresponding to this region shows distinctive helical properties when examined by circular dichroism spectroscopy. Finally, evidence is presented showing that GABP beta 1-1 and GABP beta 2-1 can heterodimerize through this carboxy-terminal domain, but neither protein can heterodimerize via the dimer-forming region of the bZIP protein C/EBP beta.
Tularik, Inc., South San Francisco, California 94080.
F C
FC
de la Brousse
E H
EH
Birkenmeier
D S
DS
King
L B
LB
Rowe
S L
SL
McKnight
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D001704:Biopolymers; D015153:Blotting, Western; D002874:Chromosome Mapping; D002942:Circular Dichroism; D004268:DNA-Binding Proteins / Q000032:analysis / Q000737:chemistry* / Q000235:genetics*; D051268:GA-Binding Protein Transcription Factor; D051379:Mice; D008969:Molecular Sequence Data; D017433:Protein Structure, Secondary; D014018:Tissue Distribution; D014157:Transcription Factors / Q000032:analysis / Q000737:chemistry* / Q000235:genetics*
D001704:Biopolymers; D004268:DNA-Binding Proteins; D051268:GA-Binding Protein Transcription Factor; C495656:Gabpa protein, mouse; C495658:Gabpb1 protein, mouse; C495659:Gabpb2 protein, mouse; D014157:Transcription Factors