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Abstract
For characterization of the detailed gene structure of human muscle type carnitine palmitoyltransferase I (M-CPTI), we analyzed the 5'-upstream region of the M-CPTI transcripts. As a result, we found a cDNA clone containing a nucleotide sequence unexpected from the reported M-CPTI gene structure in the upstream region of its 5' end. Comparison of this nucleotide sequence with that of genomic DNA showed that this sequence was derived from the 3'-untranslated region of the gene encoding choline/ethanolamine kinase-beta (CK/EK-beta) located upstream of the M-CPTI gene. Southern blot analysis showed that there was no other region homologous to the CK/EK-beta gene in the whole human genome. Thus, the overlapping transcript was concluded to be produced from the functional genes of CK/EK-beta and M-CPTI. Furthermore, cDNAs containing both exons of these genes were detected by the polymerase chain reaction using the cDNA of human heart M-CPTI obtained by specific reverse transcription from its 3'-untranslated region as a template. From these results, the production and organization of these overlapping transcripts are discussed.
Highlights
The nucleotide sequence(s) reported in this paper has been submitted to the GenBankTM/EBI Data Bank with accession number(s) AB029885 and AB029886
Exon M corresponds to exon 1B, whereas the nucleotide of the 5Ј end of exon U is the cytosine base shown by the symbol (#) located inside exon 1A
Comparison of the nucleotide sequences of the SLIC-polymerase chain reaction (PCR) clones obtained with that of the human M-CPTI gene (7) showed that most cDNA clones contained either exon 1A or 1B to various extents, both being linked to exon 2, as we reported previously (7)
Summary
Materials and General Methods—Human poly(A)ϩ RNAs, genomic DNA and a placental genomic DNA library were obtained from CLONTECH (Palo Alto, CA), and T4 RNA ligase and EX Taq polymerase were from TaKaRa Shuzo (Tokyo, Japan).
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