Mutations in the C-terminal domain of topoisomerase II affect meiotic function and interaction with the casein kinase 2 β subunit

• DOI: 10.1007/978-1-4419-8624-5_11
• Publication Date: 2011-02-24T03:02:08Z
• Authors: D. Leroy; G. C. Alghisi; E. Roberts; O. Filhol-Cochet; S. M. Gasser
• Keywords: Base Sequence; Molecular Sequence Data; DNA, Recombinant; Saccharomyces cerevisiae; Protein Serine-Threonine Kinases; Peptide Mapping; Meiosis; DNA Topoisomerases, Type II; Mutagenesis, Site-Directed; Electrophoresis, Gel, Two-Dimensional; Amino Acid Sequence; Phosphorylation; Casein Kinase II; Protein Binding

Topoisomerase II is a major target of the protein kinase casein kinase 2 (PK CK2) in vivo. All major phosphorylation acceptor sites in the yeast enzyme are found in the C-terminal 350aa. The acceptor sites are generally clustered such that there is more than one modified Ser or Thr within a short peptide. Mutagenesis of the predicted acceptor sites have confirmed that five of the eight predicted sites are targeted in vitro and in vivo by PK CK2. Mutation to nonphosphorylatable, neutral residues provokes at most a 10% increase in mitotic doubling time. Truncation of the enzyme leaves the enzyme catalytically active, but slightly lengthens the doubling time during mitotic growth and impedes progress through meiosis. Since this could reflect the loss of interaction with an important ligand, we have examined whether the C-terminal domain of the yeast enzyme mediates interaction with the regulatory beta subunit of PK CK2, which was previously reported to bind topoisomerase II. We find that point mutation of the phospho-acceptor sites does not abrogate the interaction with a small region of PK CK2beta, while truncation at aa1276 or aa1236 does. The site of interaction within PK CK2beta does not coincide with the highly negatively charged spermine binding site.