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The pentafunctional FAS1 gene of yeast: its nucleotide sequence and order of the catalytic domains

Summary

FAS1, the structural gene of the pentafunctional fatty acid synthetase subunit β in Saccharomyces cerevisiae has been sequenced. Its reading frame represents an intronfree nucleotide sequence of 5,535 base pairs, corresponding to a protein of 1,845 amino acids with a molecular weight of 205,130 daltons. In addition to the coding sequence, 1,468 base pairs of its 5′-flanking region were determined. S1 nuclease mapping revealed two transcriptional initiation sites, 5 and 36 base pairs upstream of the translational start codon. Within the flanking sequences two TATATAAA boxes, several A-rich and T-rich blocks and a TAG-...TATGTT...TATGTT...TTT sequence were found and are discussed as transcriptional initiation and termination signals, respectively. The order of catalytic domains in the cluster gene was established by complementation of defined fas1 mutants with overlapping FAS1 subclones. Acetyl transferase (amino acids 1–468) is located proximal to the N-terminus of subunit β, followed by the enoyl reductase (amino acids 480–858), the dehydratase (amino acids 1,134–1,615) and the malonyl/palmityl transferase (amino acids 1,616–1,845) domains. One major inter-domain region of about 276 amino acids with so far unknown function was found between the enoyl reductase and dehydratase domains. The substrate-binding serine residues of acetyl, malonyl and palmityl transferases were identified within the corresponding domains. Significant sequence homologies exist between the acyl transferase active sites of yeast and animal fatty acid synthetases. Similarly, a putative sequence of the enoyl reductase active site was identified.

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Correspondence to Michael Schweizer.

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Communicated by C.P. Hollenberg

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Schweizer, M., Roberts, L.M., Höltke, H. et al. The pentafunctional FAS1 gene of yeast: its nucleotide sequence and order of the catalytic domains. Mol Gen Genet 203, 479–486 (1986). https://doi.org/10.1007/BF00422073

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Key words

  • Multifunctional enzyme
  • Cluster gene
  • Nucleotide sequence
  • Yeast transformation