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Identification of Mutations in Protein Kinase CKIIβ Subunit That Affect Its Binding to Ribosomal Protein L41 and Homodimerization

  • Ahn, Bong-Hyun (Institutions Department of Biochemistry, College of Natural Sciences, Kyungpook National University) ;
  • Lee, Ji-Hoon (Institutions Department of Biochemistry, College of Natural Sciences, Kyungpook National University) ;
  • Bae, Young-Seuk (Institutions Department of Biochemistry, College of Natural Sciences, Kyungpook National University)
  • Received : 2003.01.30
  • Accepted : 2003.02.14
  • Published : 2003.07.31

Abstract

Protein kinase CKII is composed of two catalytic ($\alpha$ or $\alpha$') subunits and two regulatory ($\beta$) subunits. The $CKII{\beta}$ subunit is thought to mediate the tetramer formation and interact with other target proteins. However, its physiological function remains obscure. In this study, point mutants of $CKII{\beta}$ that are defective for the L41 binding were isolated by using the reverse two-hybrid system. A sequence analysis of the point mutants revealed that Asp-26, Met-52, and Met-78 of $CKII{\beta}$ are critical for L41 binding; Asn-67 (and/or Lys-139) and Met-52 are important for $CKII{\beta}$ homodimerization. Two point mutants, R75 and R83, of $CKII{\beta}$ interacted with L5, topoisomerase $II{\beta}$, and CKBBP1/SAG, but not with the wild-type $CKII{\beta}$. This indicates that $CKII{\beta}$ homodimerization is not a prerequisite for its binding to target proteins. These $CKII{\beta}$ point mutants may be useful in exploring the biochemical physiological functions of $CKII{\beta}$.

Keywords

Protein kinase CKII;Protein-protein interaction;Random PCR mutagenesis;Reverse two-hybrid system

Acknowledgement

Supported by : Korea Science & Engineering Foundation

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