A novel splicing variant encoding putative catalytic alpha subunit of maize protein kinase CK2.
A cDNA highly homologous to the known catalytic alpha subunit of protein kinase CK2 was cloned from maize (Zea mays). It was designated ZmCK2alpha-4 (accession no. AAF76187). Sequence analysis shows that ZmCK2alpha-4 and the previously identified ZmCK2alpha-1 (accession no. X61387) are transcribed from the same gene, ZmPKCK2AL (accession no. Y11649), but at different levels in various maize organs and at different stages of development. The cDNA encoding ZmCK2alpha-4 has three potential translation initiation sites. The three putative variants of ZmCK2alpha-4 were expressed in Escherichia coli as GST-fusion proteins and purified from bacterial extracts. In contrast to the previously characterized ZmCK2alphas, the obtained GST:ZmCK2alpha-4 proteins were catalytically inactive as monomers or in the presence of equimolar amounts of the human CK2beta. However, GST:ZmCK2alpha-4 did phosphorylate casein in the presence of a large excess of the beta subunit. The activity of ZmCK2alpha-4 toward casein could also be stimulated by increasing ATP concentration. Modeling studies have shown that there is no interaction between the N-terminal segment of ZmCK2alpha-4 and the activation loop responsible for constitutive catalytic activity of CK2alpha. Preliminary results suggest that ZmCK2alpha-4 may function as a negative regulator of other CK2s, and at certain circumstances as a holoenzyme which catalytic activity is stimulated by specific regulatory subunit(s).
Lebska
Maja
M
Institute of Biochemistry and Biophysics, Polish Academy of Sciences, Pawińskiego 5a, 02-106 Warsaw, Poland.
Szczegielniak
Jadwiga
J
Dobrowolska
Grazyna
G
Cozza
Giorgio
G
Moro
Stefano
S
Muszyńska
Grazyna
G
D000595:Amino Acid Sequence; D047390:Casein Kinase II / Q000235:genetics* / Q000378:metabolism; D002364:Caseins / Q000378:metabolism; D020134:Catalytic Domain; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000235:genetics; D007527:Isoenzymes / Q000235:genetics / Q000378:metabolism; D008958:Models, Molecular; D008969:Molecular Sequence Data; D010766:Phosphorylation; D018749:RNA, Plant / Q000235:genetics; D016415:Sequence Alignment; D003313:Zea mays / Q000201:enzymology / Q000235:genetics*
D002364:Caseins; D018076:DNA, Complementary; D007527:Isoenzymes; D018749:RNA, Plant; D047390:Casein Kinase II