Molecular shape and prominent role of β-strand swapping in organization of dUTPase oligomers

Abstract 

Most dUTP pyrophosphatases (dUTPases) are homotrimers with interfaces formed between subunit surfaces, in the central channel, and by C-terminal β-strand swapping. Analysis of intersubunit interactions reveals an important cohesive role for the C-terminus. This is reflected in the crystal structure of fruitfly dUTPase displaying a dimeric organization in crystals grown in alcohol solution, where only β-strand swapping interactions between subunits are retained from the usual trimer structure. Mutations of a suggested hinge proline destabilize human and Escherichia coli dUTPases without preventing trimeric organization. Trimer formation was, however, prevented in the human enzyme by truncating the C-terminus before the swapping arm. The molecular shape of full-length enzymes in solution reveals the localization and variation in flexibility of N- and C-terminal segments.

Structured summary

MINT-6946477:dUTPase (uniprotkb:Q9V3I1) and dUTPase (uniprotkb:Q9V3I1) bind (MI:0407) by X-ray crystallography (MI:0114)

Abbreviations: E. coli, Escherichia coli, dUTP, 2′-deoxyuridine triphosphate, dUTPase, dUTP pyrophosphatase, MPD, 2-methyl-2,4-pentanediol, DTT, 1,4-dithiothreitol, PMSF, phenylmethylsulfonyl fluoride, SAXS, small angle X-ray scattering, CD, circular dichroism, DSC, differential scanning calorimetry.

Keywords: dUTP pyrophosphatase, Oligomerization, Subunit interactions, β-Strand swapping, Hinge proline