Dissecting subdomains involved in multiple functions of the CK2beta subunit.
We have characterized several subdomains of the beta subunit of protein kinase CK2. The N-terminal half of the protein exhibits a pseudo-substrate segment in tandem with a polyamine binding domain responsible for the activation of the kinase by these polybasic compounds. Study of the chemical features of this polyamine binding site showed that polyamine analogs exhibiting the highest affinity for CK2 are the best CK2 activators. Mutational analysis disclosed that glutamic residues lying in the polyacidic region of the CK2beta subunit are involved in the interaction with polyamine molecules and allowed the delineation of an autonomous binding domain. Furthermore, this regulatory domain was shown to mediate the association of CK2 with plasma membrane. The C-terminal domain of the CK2beta subunit plays a role in the oligomerization of the kinase since it was observed that a truncated form of this subunit lacking its 33-last amino acids was incompetent for the assembly of polymeric forms of CK2. Altogether, our results support the notion that the beta subunit of CK2 is a modular protein made by the association of interdependent domains that are involved in its multiple functions.
Leroy
D
D
Laboratoire de Biochimie des Régulations Cellulaires Endocrines, INSERM U244, DBMS, CEA Grenoble, France.
Filhol
O
O
Quintaine
N
N
Sarrouilhe
D
D
Loue-Mackenbach
P
P
Chambaz
E M
EM
Cochet
C
C
D000345:Affinity Labels; D000595:Amino Acid Sequence; D000818:Animals; D001665:Binding Sites; D001704:Biopolymers; D047390:Casein Kinase II; D002462:Cell Membrane / Q000201:enzymology; D004789:Enzyme Activation; D006801:Humans; D008099:Liver / Q000201:enzymology; D008297:Male; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D011073:Polyamines / Q000378:metabolism; D017346:Protein Serine-Threonine Kinases / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D051381:Rats; D017208:Rats, Wistar; D017386:Sequence Homology, Amino Acid; D013379:Substrate Specificity
D000345:Affinity Labels; D001704:Biopolymers; D011073:Polyamines; D047390:Casein Kinase II; D017346:Protein Serine-Threonine Kinases