A second form (beta isoform) of nucleoside diphosphate kinase from rat. Isolation and characterization of complementary and genomic DNA and expression.
Complementary and genomic clones for a second form (the beta isoform) of rat nucleoside diphosphate kinase were isolated. Structural studies revealed that nucleotide and deduced amino acid sequences of the beta isoform were quite similar to those of the alpha isoform (identities were 82 and 89%, respectively), which were delineated in our previous study (Kimura, N., Shimada, N., Nomura, K., and Watanabe, K. (1990) J. Biol. Chem. 265, 15744-15749). The gene encoding the beta isoform, covering 10 kilobases and comprising five exons, was located in tandem in the immediate vicinity (at a 3-kilobase distance) of the 5' upstream of the alpha isoform gene which was recently reported from this laboratory (Ishikawa, N., Shimada, N., Munakata, Y., Watanabe, K., and Kimura, N. (1992) J. Biol. Chem. 267, 14366-14372), suggesting their generation by gene duplication. The exon-intron junctions were exactly conserved between the two genes. Southern blot analyses showed that unidentified fragments cross-reacted with the beta isoform cDNA probe besides those containing the genuine gene, and at least two of them were identified as possible processed pseudogenes. Northern and dot blot hybridization studies demonstrated that the alpha isoform was more expressed than the beta isoform in rat tissues examined except brain, from which the isoform designation was derived. These results suggest independent expression and specific roles of these isoforms in the cell. Comparative studies between rat and human isoforms indicate that the isoforms could have differentiated before the two species evolutionally diverged.
Department of Molecular Biology, Tokyo Metropolitan Institute of Gerontology, Japan.
N
N
Shimada
N
N
Ishikawa
Y
Y
Munakata
T
T
Toda
K
K
Watanabe
N
N
Kimura
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D003001:Cloning, Molecular; D004247:DNA / Q000235:genetics; D015870:Gene Expression; D005796:Genes; D007527:Isoenzymes / Q000235:genetics; D008969:Molecular Sequence Data; D009701:Nucleoside-Diphosphate Kinase / Q000235:genetics*; D009838:Oligodeoxyribonucleotides / Q000737:chemistry; D011544:Pseudogenes; D012333:RNA, Messenger / Q000235:genetics; D051381:Rats; D015183:Restriction Mapping; D016415:Sequence Alignment
D007527:Isoenzymes; D009838:Oligodeoxyribonucleotides; D012333:RNA, Messenger; D004247:DNA; D009701:Nucleoside-Diphosphate Kinase