Isolation and identification of 3-methylcrotonyl coenzyme A carboxylase cDNAs and pyruvate carboxylase, and their expression in red seabream (Pagrus major) organs.
We determined complementary DNA sequences of biotin-containing (MCCC1) and non-biotin-containing (MCCC2) subunits of 3-methylcrotonyl coenzyme A carboxylase (MCCase) and pyruvate carboxylase (PCase) using reverse transcriptase polymerase chain reaction of RNA extracted from seabream skeletal muscle and liver. We determined the complete coding sequences of MCCC1 and PC and a partial coding sequence of the major part of MCCC2. Molecular sizes of MCCC1, MCCC2, and PC were 4300, 2400, and 6500 nucleotides, respectively, according to Northern blot analysis. The length of MCCC1 from cDNA sequencing was 4249 nucleotides, indicating the full-length messenger RNA sequence was obtained. Northern blot analyses showed that PC was expressed in muscle, heart, liver, and ovary, but not in spleen. MCCC1 and MCCC2 were expressed at high levels in muscle and ovary, but only trace levels in heart, spleen, and liver. MCCase appears to be particularly important in muscle and ovary, which are active in protein metabolism, while PCase is important in organs active in glycolysis, such as liver.
Abe
Shunnosuke
S
Laboratory of Molecular Cell Biology, Faculty of Agriculture, Ehime University, 3-5-7 Tarumi, Matsuyama City, 790-8566, Japan. abe@mcb.agr.ehime-u.ac.jp
Chamnan
Chhoun
C
Miyamoto
Kenichi
K
Minamino
Yasutaka
Y
Nouda
Makoto
M
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D015152:Blotting, Northern; D019735:Carbon-Carbon Ligases / Q000235:genetics* / Q000378:metabolism*; D003001:Cloning, Molecular; D016000:Cluster Analysis; D017931:DNA Primers; D018076:DNA, Complementary / Q000235:genetics; D004591:Electrophoresis, Polyacrylamide Gel; D005260:Female; D015870:Gene Expression; D008099:Liver / Q000378:metabolism; D008969:Molecular Sequence Data; D018482:Muscle, Skeletal / Q000378:metabolism; D010053:Ovary / Q000378:metabolism; D010473:Perciformes / Q000235:genetics*; D010802:Phylogeny; D011766:Pyruvate Carboxylase / Q000235:genetics* / Q000378:metabolism*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D016415:Sequence Alignment; D017422:Sequence Analysis, DNA
D017931:DNA Primers; D018076:DNA, Complementary; D019735:Carbon-Carbon Ligases; D011766:Pyruvate Carboxylase; C021573:methylcrotonoyl-CoA carboxylase