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. 1996 Jan;178(1):121–129. doi: 10.1128/jb.178.1.121-129.1996

The sea pansy Renilla reniformis luciferase serves as a sensitive bioluminescent reporter for differential gene expression in Candida albicans.

T Srikantha 1, A Klapach 1, W W Lorenz 1, L K Tsai 1, L A Laughlin 1, J A Gorman 1, D R Soll 1
PMCID: PMC177628  PMID: 8550405

Abstract

The infectious yeast Candida albicans progresses through two developmental programs which involve differential gene expression, the bud-hypha transition and high-frequency phenotypic switching. To understand how differentially expressed genes are regulated in this organism, the promoters of phase-specific genes must be functionally characterized, and a bioluminescent reporter system would facilitate such characterization. However, C. albicans has adopted a nontraditional codon strategy that involves a tRNA with a CAG anticodon to decode the codon CUG as serine rather than leucine. Since the luciferase gene of the sea pansy Renilla reinformis contains no CUGs, we have used it to develop a highly sensitive bioluminescent reporter system for C. albicans. When fused to the galactose-inducible promoter of GAL1, luciferase activity is inducible; when fused to the constitutive EF1 alpha 2 promoter, luciferase activity is constitutive; and when fused to the promoter of the white-phase-specific gene WH11 or the opaque-phase-specific gene OP4, luciferase activity is phase specific. The Renilla luciferase system can, therefore, be used as a bioluminescent reporter to analyze the strength and developmental regulation of C. albicans promoters.

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Selected References

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  1. Bedell G. W., Soll D. R. Effects of low concentrations of zinc on the growth and dimorphism of Candida albicans: evidence for zinc-resistant and -sensitive pathways for mycelium formation. Infect Immun. 1979 Oct;26(1):348–354. doi: 10.1128/iai.26.1.348-354.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Bergen M. S., Voss E., Soll D. R. Switching at the cellular level in the white-opaque transition of Candida albicans. J Gen Microbiol. 1990 Oct;136(10):1925–1936. doi: 10.1099/00221287-136-10-1925. [DOI] [PubMed] [Google Scholar]
  3. Birse C. E., Irwin M. Y., Fonzi W. A., Sypherd P. S. Cloning and characterization of ECE1, a gene expressed in association with cell elongation of the dimorphic pathogen Candida albicans. Infect Immun. 1993 Sep;61(9):3648–3655. doi: 10.1128/iai.61.9.3648-3655.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Cannon R. D., Jenkinson H. F., Shepherd M. G. Isolation and nucleotide sequence of an autonomously replicating sequence (ARS) element functional in Candida albicans and Saccharomyces cerevisiae. Mol Gen Genet. 1990 Apr;221(2):210–218. doi: 10.1007/BF00261723. [DOI] [PubMed] [Google Scholar]
  5. Church G. M., Gilbert W. Genomic sequencing. Proc Natl Acad Sci U S A. 1984 Apr;81(7):1991–1995. doi: 10.1073/pnas.81.7.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Fonzi W. A., Irwin M. Y. Isogenic strain construction and gene mapping in Candida albicans. Genetics. 1993 Jul;134(3):717–728. doi: 10.1093/genetics/134.3.717. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Gorman J. A., Chan W., Gorman J. W. Repeated use of GAL1 for gene disruption in Candida albicans. Genetics. 1991 Sep;129(1):19–24. doi: 10.1093/genetics/129.1.19. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Hube B., Monod M., Schofield D. A., Brown A. J., Gow N. A. Expression of seven members of the gene family encoding secretory aspartyl proteinases in Candida albicans. Mol Microbiol. 1994 Oct;14(1):87–99. doi: 10.1111/j.1365-2958.1994.tb01269.x. [DOI] [PubMed] [Google Scholar]
  9. Inouye S., Noguchi M., Sakaki Y., Takagi Y., Miyata T., Iwanaga S., Miyata T., Tsuji F. I. Cloning and sequence analysis of cDNA for the luminescent protein aequorin. Proc Natl Acad Sci U S A. 1985 May;82(10):3154–3158. doi: 10.1073/pnas.82.10.3154. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Kurtz M. B., Marrinan J. Isolation of hem3 mutants from Candida albicans by sequential gene disruption. Mol Gen Genet. 1989 May;217(1):47–52. doi: 10.1007/BF00330941. [DOI] [PubMed] [Google Scholar]
  11. Leuker C. E., Ernst J. F. Toxicity of a heterologous leucyl-tRNA (anticodon CAG) in the pathogen Candida albicans: in vivo evidence for non-standard decoding of CUG codons. Mol Gen Genet. 1994 Oct 28;245(2):212–217. doi: 10.1007/BF00283269. [DOI] [PubMed] [Google Scholar]
  12. Lorenz W. W., McCann R. O., Longiaru M., Cormier M. J. Isolation and expression of a cDNA encoding Renilla reniformis luciferase. Proc Natl Acad Sci U S A. 1991 May 15;88(10):4438–4442. doi: 10.1073/pnas.88.10.4438. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Masuda T., Tatsumi H., Nakano E. Cloning and sequence analysis of cDNA for luciferase of a Japanese firefly, Luciola cruciata. Gene. 1989 Apr 30;77(2):265–270. doi: 10.1016/0378-1119(89)90074-7. [DOI] [PubMed] [Google Scholar]
  14. Matthews J. C., Hori K., Cormier M. J. Purification and properties of Renilla reniformis luciferase. Biochemistry. 1977 Jan 11;16(1):85–91. doi: 10.1021/bi00620a014. [DOI] [PubMed] [Google Scholar]
  15. Morrow B., Srikantha T., Anderson J., Soll D. R. Coordinate regulation of two opaque-phase-specific genes during white-opaque switching in Candida albicans. Infect Immun. 1993 May;61(5):1823–1828. doi: 10.1128/iai.61.5.1823-1828.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Morrow B., Srikantha T., Soll D. R. Transcription of the gene for a pepsinogen, PEP1, is regulated by white-opaque switching in Candida albicans. Mol Cell Biol. 1992 Jul;12(7):2997–3005. doi: 10.1128/mcb.12.7.2997. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Ohama T., Suzuki T., Mori M., Osawa S., Ueda T., Watanabe K., Nakase T. Non-universal decoding of the leucine codon CUG in several Candida species. Nucleic Acids Res. 1993 Aug 25;21(17):4039–4045. doi: 10.1093/nar/21.17.4039. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Prasher D. C., Eckenrode V. K., Ward W. W., Prendergast F. G., Cormier M. J. Primary structure of the Aequorea victoria green-fluorescent protein. Gene. 1992 Feb 15;111(2):229–233. doi: 10.1016/0378-1119(92)90691-h. [DOI] [PubMed] [Google Scholar]
  19. Rikkerink E. H., Magee B. B., Magee P. T. Opaque-white phenotype transition: a programmed morphological transition in Candida albicans. J Bacteriol. 1988 Feb;170(2):895–899. doi: 10.1128/jb.170.2.895-899.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Santos M. A., Keith G., Tuite M. F. Non-standard translational events in Candida albicans mediated by an unusual seryl-tRNA with a 5'-CAG-3' (leucine) anticodon. EMBO J. 1993 Feb;12(2):607–616. doi: 10.1002/j.1460-2075.1993.tb05693.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Santos M., Colthurst D. R., Wills N., McLaughlin C. S., Tuite M. F. Efficient translation of the UAG termination codon in Candida species. Curr Genet. 1990 Jun;17(6):487–491. doi: 10.1007/BF00313076. [DOI] [PubMed] [Google Scholar]
  22. Saporito-Irwin S. M., Birse C. E., Sypherd P. S., Fonzi W. A. PHR1, a pH-regulated gene of Candida albicans, is required for morphogenesis. Mol Cell Biol. 1995 Feb;15(2):601–613. doi: 10.1128/mcb.15.2.601. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Slutsky B., Staebell M., Anderson J., Risen L., Pfaller M., Soll D. R. "White-opaque transition": a second high-frequency switching system in Candida albicans. J Bacteriol. 1987 Jan;169(1):189–197. doi: 10.1128/jb.169.1.189-197.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Soll D. R. High-frequency switching in Candida albicans. Clin Microbiol Rev. 1992 Apr;5(2):183–203. doi: 10.1128/cmr.5.2.183. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Soll D. R. The regulation of cellular differentiation in the dimorphic yeast Candida albicans. Bioessays. 1986 Jul;5(1):5–11. doi: 10.1002/bies.950050103. [DOI] [PubMed] [Google Scholar]
  26. Srikantha T., Chandrasekhar A., Soll D. R. Functional analysis of the promoter of the phase-specific WH11 gene of Candida albicans. Mol Cell Biol. 1995 Mar;15(3):1797–1805. doi: 10.1128/mcb.15.3.1797. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Srikantha T., Morrow B., Schröppel K., Soll D. R. The frequency of integrative transformation at phase-specific genes of Candida albicans correlates with their transcriptional state. Mol Gen Genet. 1995 Feb 6;246(3):342–352. doi: 10.1007/BF00288607. [DOI] [PubMed] [Google Scholar]
  28. Srikantha T., Soll D. R. A white-specific gene in the white-opaque switching system of Candida albicans. Gene. 1993 Sep 6;131(1):53–60. doi: 10.1016/0378-1119(93)90668-s. [DOI] [PubMed] [Google Scholar]
  29. Stone R. L., Matarese V., Magee B. B., Magee P. T., Bernlohr D. A. Cloning, sequencing and chromosomal assignment of a gene from Saccharomyces cerevisiae which is negatively regulated by glucose and positively by lipids. Gene. 1990 Dec 15;96(2):171–176. doi: 10.1016/0378-1119(90)90249-q. [DOI] [PubMed] [Google Scholar]
  30. Sundstrom P., Smith D., Sypherd P. S. Sequence analysis and expression of the two genes for elongation factor 1 alpha from the dimorphic yeast Candida albicans. J Bacteriol. 1990 Apr;172(4):2036–2045. doi: 10.1128/jb.172.4.2036-2045.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Suzuki T., Ueda T., Ohama T., Osawa S., Watanabe K. The gene for serine tRNA having anticodon sequence CAG in a pathogenic yeast, Candida albicans. Nucleic Acids Res. 1993 Jan 25;21(2):356–356. doi: 10.1093/nar/21.2.356. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Tatsumi H., Kajiyama N., Nakano E. Molecular cloning and expression in Escherichia coli of a cDNA clone encoding luciferase of a firefly, Luciola lateralis. Biochim Biophys Acta. 1992 Jun 15;1131(2):161–165. doi: 10.1016/0167-4781(92)90071-7. [DOI] [PubMed] [Google Scholar]
  33. White T. C., Andrews L. E., Maltby D., Agabian N. The "universal" leucine codon CTG in the secreted aspartyl proteinase 1 (SAP1) gene of Candida albicans encodes a serine in vivo. J Bacteriol. 1995 May;177(10):2953–2955. doi: 10.1128/jb.177.10.2953-2955.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. White T. C., Miyasaki S. H., Agabian N. Three distinct secreted aspartyl proteinases in Candida albicans. J Bacteriol. 1993 Oct;175(19):6126–6133. doi: 10.1128/jb.175.19.6126-6133.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Wick R. A. Photon counting imaging: applications in biomedical research. Biotechniques. 1989 Mar;7(3):262–269. [PubMed] [Google Scholar]
  36. de Wet J. R., Wood K. V., DeLuca M., Helinski D. R., Subramani S. Firefly luciferase gene: structure and expression in mammalian cells. Mol Cell Biol. 1987 Feb;7(2):725–737. doi: 10.1128/mcb.7.2.725. [DOI] [PMC free article] [PubMed] [Google Scholar]

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