dUTPase and nucleocapsid polypeptides of the Mason-Pfizer monkey virus form a fusion protein in the virion with homotrimeric organization and low catalytic efficiency

Orsolya Barabás, Michaela Rumlová, A. Erdei, Veronika Pongrácz, Iva Pichová, B. Vértessy

Research output: Contribution to journalArticle

17 Citations (Scopus)

Abstract

Betaretroviruses encode dUTPase, an essential factor in DNA metabolism and repair, in the pro open reading frame located between gag and pol. Ribosomal frameshifts during expression of retroviral proteins provide a unique possibility for covalent joining of nucleocapsid (NC) and dUTPase within Gag-Pro polyproteins. By developing an antibody against the prototype betaretrovirus Mason-Pfizer monkey virus dUTPase, we demonstrate that i) the NC-dUTPase fusion protein exists both within the virions and infected cells providing the only form of dUTPase, and ii) the retroviral protease does not cleave NC-dUTPase either in the virion or in vitro. We show that recombinant betaretroviral NC-dUTPase and dUTPase are both inefficient catalysts compared with all other dUTPases. Dynamic light scattering and gel filtration confirm that the homotrimeric organization, common among dUTPases, is retained in the NC-dUT-Pase fusion protein. The betaretroviral dUTPase has been crystallized and single crystals contain homotrimers. Oligonucleotide and Zn2+ binding is well retained in the fusion protein, which is the first example of acquisition of a functional nucleic acid binding module by the DNA repair factor dUTPase. Binding of the hexanucleotide ACTGCC or the octanucleotide (TG)4 to NC-dUT-Pase modulates enzymatic function, indicating that the low catalytic activity may be compensated by adequate localization.

Original languageEnglish
Pages (from-to)38803-38812
Number of pages10
JournalJournal of Biological Chemistry
Volume278
Issue number40
DOIs
Publication statusPublished - Oct 3 2003

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Mason-Pfizer monkey virus
Nucleocapsid
Viruses
Virion
Fusion reactions
Peptides
Proteins
Betaretrovirus
DNA Repair
Repair
Ribosomal Frameshifting
dUTP pyrophosphatase
gag Gene Products
Polyproteins
DNA
Dynamic light scattering
Metabolism
Oligonucleotides
Joining
Nucleic Acids

ASJC Scopus subject areas

  • Biochemistry

Cite this

dUTPase and nucleocapsid polypeptides of the Mason-Pfizer monkey virus form a fusion protein in the virion with homotrimeric organization and low catalytic efficiency. / Barabás, Orsolya; Rumlová, Michaela; Erdei, A.; Pongrácz, Veronika; Pichová, Iva; Vértessy, B.

In: Journal of Biological Chemistry, Vol. 278, No. 40, 03.10.2003, p. 38803-38812.

Research output: Contribution to journalArticle

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