Plant and Cell Physiology, 1995, Vol. 36, No. 5 779-787
© 1995
Genomic Organization of 251 kDa Acetyl-CoA Carboxylase Genes in Arabidopsis: Tandem Gene Duplication has Made Two Differentially Expressed Isozymes
1Mitsui Plant Biotechnology Research Institute TCID-21, Sengen 2-1-6, Tsukuba, 305 Japan
2Department of Botany, Iowa State University Ames, Iowa 50011, U.S.A.
3Department of Biochemistry and Biophysics, Iowa State University Ames, Iowa 50011, U.S.A.
Acetyl-CoA carboxylase (ACCase) catalyzes the carboxylation of acetyl-CoA, forming malonyl-CoA a key intermediate in the biosynthesis of fatty acids and a variety of secondary metabolites. Based upon amino acid sequences conserved among rat, chicken, and E. coli ACCases, PCR-primers were used to amplify a genomic fragment which codes for an ACCase of Arabidopsis. The resulting fragment was used for isolation of genomic and cDNA clones. We have determined the complete cDNA sequence coding for an Arabidopsis ACCase consists of 2,254 amino acids with the molecular mass of 251 kDa. This enzyme contains no recognizable plastid transit-peptide sequence. Therefore, this ACCase is presumably the cytosolic isozyme. Southern analysis indicates that there are two ACCase genes in the Arabidopsis genome. Surprisingly, the results of RFLP analysis and physical mapping of the isolated genomic clones demonstrate that these two genes, acc1 and acc2, are contiguously located within a 25-kbp genomic region near the middle of chromosome 1. Both genes are transcriptionally active, as transcripts from each gene were detected by reverse transcription-PCR analysis using gene-specific primers. The acc1 and acc2 transcripts accumulate in leaves and seedlings but only the acc1 transcript accumulates in developing siliques, unexpectedly. The differences in the expression patterns may be indicative of the differential role of the two genes.
(Received December 27, 1994; Accepted April 11, 1995)
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