The genomic structure of two protein kinase CK2alpha genes of Xenopus laevis and features of the putative promoter region.
Protein kinase CK2 is an enzyme that is ubiquitous in eukaryotes. This enzyme, composed of catalytic (alpha and alpha') and regulatory (beta) subunits, is responsible for the phosphorylation of a large number of proteins and is implicated in cell division. Genomic clones coding for the CK2alpha subunit of Xenopus laevis have been isolated. Initial restriction enzyme profiles and subsequent PCR analysis and DNA sequencing indicated that these genomic clones correspond to two different genes. The two genes are highly homologous in the regions of the coding sequence (only 3 amino acid differences) but differ considerable in their intron sequences and lengths. Gene 1 corresponds to the cDNA of XlCK2alpha which had been previously isolated and described. The genomic clone for this gene was truncated. Gene 2 contains the entire coding region for CK2alpha subunit as well as a fragment of 6.4 kb of the 5' upstream region. The exon/intron boundaries of both genes obey the GT/AG rule with the exception of intron V where the less common GC/AG is seen. Comparison of the size of ten coding exons and sites where these are interrupted by introns shows strong conservation with respect to the human CK2alpha gene. RT-PCR analysis of mRNAs from X. laevis ovary, oocytes and early embryos using a specific primer for gene 2 demonstrated that this gene is expressed in these tissues and cells. Analysis of transcription start sites using 5'RACE and RNA from stage VI oocytes demonstrated that there are multiple start sites in the XlCK2alpha mRNA. It was also seen that a noncoding exon 1 is present 4 kb upstream of the translation start site and that alternate splicing occurs in gene 2 to give exon 1 of different lengths. Sequencing of the entire upstream genomic region of gene 2 revealed that there are regions of homology to the sequence of exon 1 of the human CK2alpha gene. Other sequences with consensus to transcription factor binding sites that are seen in the promoter region of human CK2alpha are also found in the X. laevis CK2alpha gene 2. These sites include Ets1, E2F, CCAAT and GC rich regions. No canonical TATA motif is observed.
Wilhelm
V
V
Programa de Biología Celular y Molecular, Instituto de Ciencias Biomédicas, Facultad de Medicina, Universidad de Chile, Santiago.
Neckelman
G
G
Allende
J E
JE
Allende
C C
CC
D020816:Amino Acid Motifs; D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D001665:Binding Sites; D047390:Casein Kinase II; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000378:metabolism; D005091:Exons; D006801:Humans; D007438:Introns; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction; D011401:Promoter Regions, Genetic; D017346:Protein Serine-Threonine Kinases / Q000235:genetics*; D017434:Protein Structure, Tertiary; D050784:Proto-Oncogene Protein c-ets-1; D011518:Proto-Oncogene Proteins / Q000378:metabolism; D050783:Proto-Oncogene Proteins c-ets; D012333:RNA, Messenger / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D017386:Sequence Homology, Amino Acid; D012689:Sequence Homology, Nucleic Acid; D014157:Transcription Factors / Q000378:metabolism; D014158:Transcription, Genetic; D014982:Xenopus laevis / Q000235:genetics* / Q000378:metabolism
D018076:DNA, Complementary; C494041:ETS1 protein, human; D050784:Proto-Oncogene Protein c-ets-1; D011518:Proto-Oncogene Proteins; D050783:Proto-Oncogene Proteins c-ets; D012333:RNA, Messenger; D014157:Transcription Factors; D047390:Casein Kinase II; D017346:Protein Serine-Threonine Kinases