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Nucleic Acids Research, 1985, Vol. 13, No. 12 4365-4378
© 1985

Articles

Molecular cloning and the nudeotide sequence of cDNA to mRNA for Don-nearonal enolase ({alpha}{alpha} enolase) of rat brain and liver

Kenji Sakimura, Etsuko Kushiya, Masuo Obinata* and Yasuo Takahashi

*Faculty of Pharmaceutical Science, Tokyo University Hongo 7-3-1, Tokyo 113, Japan Department of Neurophannacology, Brain Research Institute, Niigata University Asahimachi 1, Niigata 951

Received March 12, 1985. Revised May 21, 1985. Accepted May 21, 1985.

The nucleotide sequence for {alpha}{alpha} enolase (non-neuronal enolase: NNE) of rat brain and liver was determined from recombinant cDNA clones. The sequence was composed of 1722 bp which included the 1299 bp of the complete coding region, the 108 bp of the 5'-noncoding region and the 312 bp of the 3'-noncoding region containing a polyadenylation signal. In addition, the poly(A) tail was also found. A potential ribosome-binding site was located 30 nucleotides upstream to the initiation codon in the 5'-noncoding region. The amino acid sequence deduced from the nucleotide sequence was 433 amino acids in length and showed very high homology (82%) to the amino acid sequence of {gamma}{gamma} enolase (neuron-specific enolase: NSE), although the nucleotide sequence showed slightly lower homology (75%). The size of NNE mRNA was approximately 1800 bases by Northern transfer analysis and much shorter than that of NSE mRNA (2400 bases) indicating a short 3'-noncoding region. A dot-blot hybridization and Northern transfer analysis of cytoplasmic RNA from the developing rat brains using a labeled 3'-noncoding region of cDNA (no homology between NSE and NNE) showed a decrease of NNE mRNA at around 10 postnatal days and then a gradual increase to adult age without changes of mRNA size. Liver mRNA did not show any significant change during development


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