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. 2007 May 29;404(Pt 3):459–466. doi: 10.1042/BJ20061611

Figure 5. Actin-related motif in the Neh5 domain is conserved among CNC family transcription factors.

Figure 5

(A) Alanine-scanning mutation analysis of the Nrf2 Neh5 domain. The mutations (M1–M15) were introduced into GBD–Neh5 by site-directed mutagenesis. Gal4–TATA–Luc reporter (20 ng) and GBD-Neh5 [30 ng; WT (wild-type)] or GBD–Neh5 M1 to GBD–Neh5M15 fusion protein expression constructs were co-transfected into QT6 cells. Luciferase assay was as described in (Figure 3B). *P<0.05, **P<0.01 relative to the GBD–Neh5 (WT) (Dunnett's test). (B) Immunoblot analysis of Neh5 mutants. QT6 whole cell lysates transfected with GBD–Neh5 (lane 1) or GBD–Neh5M1 to GBD–Neh5M15 (lanes 2–16) expression constructs were subjected to immunoblotting using anti-GBD or anti-β-Gal antibodies. (C) An alignment of the actin-related motif in the Neh5 domain among CNC family transcription factors. Amino acid residues identical among at least three proteins are shaded in grey. Acidic residues and hydrophobic residues conserved between CNC proteins and ARP1 are indicated by asterisks and closed circles respectively. The consensus sequence of actin-related motif is shown below.