Functional analysis of the promoter of the phase-specific WH11 gene of Candida albicans

Srikantha Thyagarajan, A. Chandrasekhar, D. R. Soll

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Abstract

Candida albicans WO-1 switches spontaneously, frequently, and reversibly between a hemispherical white and a fiat gray (opaque) colony-forming phenotype. This transition affects a number of morphological and physiological parameters and involves the activation and deactivation of phase-specific genes. The WH11 gene is transcribed in the white but not the opaque phase. A chimeric WH11-firefly luciferase gene containing the 5' upstream region of WH11 was demonstrated to be under phase regulation regardless of the site of integration, and a series of promoter deletion constructs was used to delineate two white-phase-specific transcription activation domains. Gel retardation experiments with the individual distal or proximal domain and white-phase or opaque-phase protein extract demonstrated the formation of one distal white-phase-specific complex and two proximal white-phase-specific complexes. Specific subfragments were tested for their ability to compete with the entire domain in the formation of complexes with white-phase protein extract in order to map the proximal domain sequence involved in white-phase-specific complex formation. Our results indicate that white-phase-specific transcription of WH11 is positively regulated by trans- acting factors interacting with two cis-acting activation sequences in the WH11 promoter.

Original languageEnglish
Pages (from-to)1797-1805
Number of pages9
JournalMolecular and Cellular Biology
Volume15
Issue number3
StatePublished - 1 Jan 1995

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Candida albicans
Genes
Firefly Luciferases
Trans-Activators
Transcriptional Activation
Proteins
Gels
Phenotype

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title = "Functional analysis of the promoter of the phase-specific WH11 gene of Candida albicans",
abstract = "Candida albicans WO-1 switches spontaneously, frequently, and reversibly between a hemispherical white and a fiat gray (opaque) colony-forming phenotype. This transition affects a number of morphological and physiological parameters and involves the activation and deactivation of phase-specific genes. The WH11 gene is transcribed in the white but not the opaque phase. A chimeric WH11-firefly luciferase gene containing the 5' upstream region of WH11 was demonstrated to be under phase regulation regardless of the site of integration, and a series of promoter deletion constructs was used to delineate two white-phase-specific transcription activation domains. Gel retardation experiments with the individual distal or proximal domain and white-phase or opaque-phase protein extract demonstrated the formation of one distal white-phase-specific complex and two proximal white-phase-specific complexes. Specific subfragments were tested for their ability to compete with the entire domain in the formation of complexes with white-phase protein extract in order to map the proximal domain sequence involved in white-phase-specific complex formation. Our results indicate that white-phase-specific transcription of WH11 is positively regulated by trans- acting factors interacting with two cis-acting activation sequences in the WH11 promoter.",
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Functional analysis of the promoter of the phase-specific WH11 gene of Candida albicans. / Thyagarajan, Srikantha; Chandrasekhar, A.; Soll, D. R.

In: Molecular and Cellular Biology, Vol. 15, No. 3, 01.01.1995, p. 1797-1805.

Research output: Contribution to journalArticleResearchpeer-review

TY - JOUR

T1 - Functional analysis of the promoter of the phase-specific WH11 gene of Candida albicans

AU - Thyagarajan, Srikantha

AU - Chandrasekhar, A.

AU - Soll, D. R.

PY - 1995/1/1

Y1 - 1995/1/1

N2 - Candida albicans WO-1 switches spontaneously, frequently, and reversibly between a hemispherical white and a fiat gray (opaque) colony-forming phenotype. This transition affects a number of morphological and physiological parameters and involves the activation and deactivation of phase-specific genes. The WH11 gene is transcribed in the white but not the opaque phase. A chimeric WH11-firefly luciferase gene containing the 5' upstream region of WH11 was demonstrated to be under phase regulation regardless of the site of integration, and a series of promoter deletion constructs was used to delineate two white-phase-specific transcription activation domains. Gel retardation experiments with the individual distal or proximal domain and white-phase or opaque-phase protein extract demonstrated the formation of one distal white-phase-specific complex and two proximal white-phase-specific complexes. Specific subfragments were tested for their ability to compete with the entire domain in the formation of complexes with white-phase protein extract in order to map the proximal domain sequence involved in white-phase-specific complex formation. Our results indicate that white-phase-specific transcription of WH11 is positively regulated by trans- acting factors interacting with two cis-acting activation sequences in the WH11 promoter.

AB - Candida albicans WO-1 switches spontaneously, frequently, and reversibly between a hemispherical white and a fiat gray (opaque) colony-forming phenotype. This transition affects a number of morphological and physiological parameters and involves the activation and deactivation of phase-specific genes. The WH11 gene is transcribed in the white but not the opaque phase. A chimeric WH11-firefly luciferase gene containing the 5' upstream region of WH11 was demonstrated to be under phase regulation regardless of the site of integration, and a series of promoter deletion constructs was used to delineate two white-phase-specific transcription activation domains. Gel retardation experiments with the individual distal or proximal domain and white-phase or opaque-phase protein extract demonstrated the formation of one distal white-phase-specific complex and two proximal white-phase-specific complexes. Specific subfragments were tested for their ability to compete with the entire domain in the formation of complexes with white-phase protein extract in order to map the proximal domain sequence involved in white-phase-specific complex formation. Our results indicate that white-phase-specific transcription of WH11 is positively regulated by trans- acting factors interacting with two cis-acting activation sequences in the WH11 promoter.

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