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Characterization of a gene in the car cluster of Fusarium fujikuroi that codes for a protein of the carotenoid oxygenase family

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Abstract

The ascomycete Fusarium fujikuroi produces carotenoids by means of the enzymes encoded by three car genes. The enzymes encoded by carRA and carB are responsible of the synthesis of β-carotene and torulene, respectively, while the product encoded by carT cleaves torulene to produce the acidic xanthophyll neurosporaxanthin. carRA and carB are found in a cluster with a third gene, carO, which codes for an opsin-like protein. However, no information is available on the sequence or chromosomal location of carT, which has been identified only by mutant analysis. Transcription of the three clustered genes is stimulated by light and by mutations in a regulatory gene, leading to overproduction of carotenoids. We have now identified a fourth gene in the car cluster, called carX, which codes for a protein similar to known carotenoid-cleaving oxygenases. carX is transcribed divergently from carRA, and exhibits the same transcriptional pattern as carRA, carB and carO. Targeted deletion of carX resulted in a phenotype characterized by a significant increase in the overall carotenoid content. In the dark, the carX mutants accumulate at least five times more carotenoids than the wild type, and exhibit partial derepression of carRA and carB transcription. The mutants also show more intense pigmentation in the light, but the increase in the carotenoid content relative to the wild type is less than twofold. Under these conditions, the mutants also show a relative increase in the amounts of phytoene and cyclic carotenoids formed, suggesting that CarRA activity is enhanced.

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Acknowledgements

We thank J. Schulte, S. Richter, C. Vallejo and L.Pérez de Camino for technical assistance. This work was supported by the German Government (DFG, grant number Tu101-7), the Spanish Government (INIA, project PB96-1336 and Ministerio de Ciencia y Tecnología, project BIO2003-01548) and the European Union (project QLK1-CT-2001-00780). This work has been carried out in compliance with the current Spanish and German laws concerning genetic research

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Correspondence to J. Avalos.

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Communicated by J. Avalos

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Thewes, S., Prado-Cabrero, A., Prado, M.M. et al. Characterization of a gene in the car cluster of Fusarium fujikuroi that codes for a protein of the carotenoid oxygenase family. Mol Genet Genomics 274, 217–228 (2005). https://doi.org/10.1007/s00438-005-0015-6

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Keywords

  • Gibberella
  • Apocarotenoid
  • Dioxygenase
  • Gene cluster