Structures of importin-α bound to the wild-type and an internal deletion mutant of the bipartite nuclear localization signal of HIF-1α

Abstract

Hypoxia-inducible factor 1 (HIF-1) is a heterodimeric transcription factor that plays an important role as a master regulator of oxygen homeostasis. The activity of HIF-1 is regulated in part by dynamic intracellular trafficking of its α subunit (HIF-1α) that can shuttle between the nucleus and cytoplasm. It has been shown that nuclear localization of HIF-1α requires a variant of classic nuclear localization signal (NLS) and that an internal deletion of the amino acid residues (residues 724–751) in the NLS almost abolish the nuclear localization. Here we report the X-ray crystal structure of the nuclear import adaptor importin-α1 bound to the wild-type HIF-1α NLS at 1.8 Å resolution and of importin-α1 bound to the Δ724-751 mutant of the HIF-1α NLS at 1.9 Å resolution. In the wild-type structure, two basic clusters in the HIF-1α NLS made extensive interactions with importin-α1 on two sites (the major site and the minor site). In the mutant structure, the NLS residues still interacted extensively with the major site on importin-α1, but the interactions with the minor site were not observed. The structural data, together with computational analyses of binding free energies, indicate that the loss of the minor-site interactions inhibit nuclear accumulation of HIF-1α.