PDBsum entry 4mz6

Protein transport

PDB id: 4mz6

Contents

Protein chain

426 a.a.

Ligands

SER-LYS-ARG-ALA-ARG-PRO-ALA

GLU-PRO-SER-LYS-ARG-ALA-ARG-PRO-ALA-GLU

Waters ×336

PDB id:

4mz6

Name:

Protein transport

Title:

Structure of importin-alpha: dutpase s11e nls mutant complex

Structure:

Deoxyuridine 5'-triphosphate nucleotidohydrolase, mitochondrial. Chain: a, c. Fragment: unp residues 97-109. Synonym: dutpase, dutp pyrophosphatase. Engineered: yes. Mutation: yes. Importin subunit alpha-1. Chain: e.

Source:

Synthetic: yes. Homo sapiens. Human. Organism_taxid: 9606. Mus musculus. Mouse. Organism_taxid: 10090. Gene: kpna2, rch1. Expressed in: escherichia coli.

Resolution:

1.88Å R-factor: 0.175 R-free: 0.195

Authors:

M.Marfori,G.Rona,B.G.Vertessy,B.Kobe

Key ref:

G.Róna et al. (2013). Phosphorylation adjacent to the nuclear localization signal of human dUTPase abolishes nuclear import: structural and mechanistic insights. Acta Crystallogr D Biol Crystallogr, 69, 2495-2505. PubMed id: 24311590 DOI: 10.1107/S0907444913023354

Date:

29-Sep-13 Release date: 13-Nov-13

Supersedes:

4fds

Protein chain

P52293  (IMA1_MOUSE) -  Importin subunit alpha-1 from Mus musculus

Seq: 529 a.a.

Struc: 426 a.a.

DOI no: 10.1107/S0907444913023354

Acta Crystallogr D Biol Crystallogr 69:2495-2505 (2013)

PubMed id: 24311590

Phosphorylation adjacent to the nuclear localization signal of human dUTPase abolishes nuclear import: structural and mechanistic insights.

G.Róna, M.Marfori, M.Borsos, I.Scheer, E.Takács, J.Tóth, F.Babos, A.Magyar, A.Erdei, Z.Bozóky, L.Buday, B.Kobe, B.G.Vértessy.

ABSTRACT

Phosphorylation adjacent to nuclear localization signals (NLSs) is involved in the regulation of nucleocytoplasmic transport. The nuclear isoform of human dUTPase, an enzyme that is essential for genomic integrity, has been shown to be phosphorylated on a serine residue (Ser11) in the vicinity of its nuclear localization signal; however, the effect of this phosphorylation is not yet known. To investigate this issue, an integrated set of structural, molecular and cell biological methods were employed. It is shown that NLS-adjacent phosphorylation of dUTPase occurs during the M phase of the cell cycle. Comparison of the cellular distribution of wild-type dUTPase with those of hyperphosphorylation- and hypophosphorylation-mimicking mutants suggests that phosphorylation at Ser11 leads to the exclusion of dUTPase from the nucleus. Isothermal titration microcalorimetry and additional independent biophysical techniques show that the interaction between dUTPase and importin-α, the karyopherin molecule responsible for `classical' NLS binding, is weakened significantly in the case of the S11E hyperphosphorylation-mimicking mutant. The structures of the importin-α-wild-type and the importin-α-hyperphosphorylation-mimicking dUTPase NLS complexes provide structural insights into the molecular details of this regulation. The data indicate that the post-translational modification of dUTPase during the cell cycle may modulate the nuclear availability of this enzyme.