RESUMO
Both, the activity as well as the expression of protein kinase CK2 is enhanced in various cancer types and in established tumour cell lines. This phenomenon is not due to an increase in the CK2 message but rather to posttranscriptional and posttranslational mechanisms. In order to get an insight into these posttranslational modifications we analyzed CK2 in prostate cancer cell lines, which differ by their hormone-sensitivity. We found that the CK2 activity is significantly higher in hormone-refractory than in hormone-sensitive cells although the amount of the catalytic alpha- and alpha'- subunits is comparable. In contrast, we detected seemingly lower amounts of the regulatory beta-subunit in the hormone-refractory cell lines, which later turned out to be an immunologically defined subclass. This subclass is realized by a phosphate group, which is attached to serine 209. The phosphorylation occurs in vivo during mitosis and is executed by the p34(cdc2)/cyclin B kinase. As this phosphorylation enhances the CK2 activity this change might well account for the higher activity of CK2 in prostate cancer cells.
Assuntos
Proteína Quinase CDC2/metabolismo , Caseína Quinase II/metabolismo , Ciclina B/metabolismo , Western Blotting , Caseína Quinase II/classificação , Caseína Quinase II/genética , Linhagem Celular Tumoral , Eletroforese em Gel de Poliacrilamida , Humanos , Masculino , Mitose , Mutação , Neoplasias Hormônio-Dependentes , Fosforilação , Neoplasias da Próstata , Subunidades Proteicas/genética , Subunidades Proteicas/metabolismo , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Serina/metabolismoRESUMO
The alpha-subunit of the casein protein kinase CK2 has been implicated in both light-regulated and circadian rhythm-controlled plant gene expression, including control of the flowering time. Two putative CK2alpha genes of perennial ryegrass (Lolium perenne L.) have been obtained from a cDNA library constructed with mRNA isolated from cold-acclimated crown tissue. The genomic organisation of the two genes was determined by Southern hybridisation analysis. Primer designs to the Lpck2a-1 and Lpck2a-2 cDNA sequences permitted the amplification of genomic products containing large intron sequences. Amplicon sequence analysis detected single nucleotide polymorphisms (SNPs) within the p150/112 reference mapping population. Validated SNPs, within diagnostic restriction enzyme sites, were used to design cleaved amplified polymorphic sequence (CAPS) assays. The Lpck2a-1 CAPS marker was assigned to perennial ryegrass linkage group (LG) 4 and the Lpck2a-2 CAPS marker was assigned to LG2. The location of the Lpck2a-1 gene locus supports the previous conclusion of conserved synteny between perennial ryegrass LG4, the Triticeae homoeologous group 5L chromosomes and the corresponding segment of rice chromosome 3. Allelic variation at the Lpck2a-1 and Lpck2a-2 gene loci was correlated with phenotypic variation for heading date and winter survival, respectively. SNP polymorphism may be used for the further study of the role of CK2alpha genes in the initiation of reproductive development and winter hardiness in grasses.
