Functional domains of the ubiquitous chromatin protein DEK.
DEK was originally described as a proto-oncogene protein and is now known to be a major component of metazoan chromatin. DEK is able to modify the structure of DNA by introducing supercoils. In order to find interaction partners and functional domains of DEK, we performed yeast two-hybrid screens and mutational analyses. Two-hybrid screening yielded C-terminal fragments of DEK, suggesting that DEK is able to multimerize. We could localize the domain to amino acids 270 to 350 and show that multimerization is dependent on phosphorylation by CK2 kinase in vitro. We also found two DNA binding domains of DEK, one on a fragment including amino acids 87 to 187 and containing the SAF-box DNA binding motif, which is located between amino acids 149 and 187. This region is sufficient to introduce supercoils into DNA. The second DNA binding domain is located between amino acids 270 and 350 and thus overlaps the multimerization domain. We show that the two DNA-interacting domains differ in their binding properties and in their abilities to respond to CK2 phosphorylation.
Kappes
Ferdinand
F
Department of Biology, University of Konstanz, Germany.
Scholten
Ingo
I
Richter
Nicole
N
Gruss
Claudia
C
Waldmann
Tanja
T
D000595:Amino Acid Sequence; D001665:Binding Sites; D047390:Casein Kinase II; D002868:Chromosomal Proteins, Non-Histone / Q000737:chemistry / Q000378:metabolism* / Q000502:physiology; D004278:DNA, Superhelical / Q000378:metabolism; D019281:Dimerization; D006367:HeLa Cells; D006801:Humans; D015513:Oncogene Proteins / Q000737:chemistry / Q000378:metabolism* / Q000502:physiology; D010766:Phosphorylation; D000075223:Poly-ADP-Ribose Binding Proteins; D017346:Protein Serine-Threonine Kinases / Q000378:metabolism; D017434:Protein Structure, Tertiary / Q000502:physiology; D000090063:Proto-Oncogene Mas; D020798:Two-Hybrid System Techniques
D002868:Chromosomal Proteins, Non-Histone; D004278:DNA, Superhelical; C084738:DEK protein, human; C000715470:MAS1 protein, human; D015513:Oncogene Proteins; D000075223:Poly-ADP-Ribose Binding Proteins; D000090063:Proto-Oncogene Mas; D047390:Casein Kinase II; D017346:Protein Serine-Threonine Kinases