Characterization of ABG1, a novel and essential gene of Candida albicans coding for a vacuolar protein involved in cytokinesis an dhyphal branching

V. Veses, M. Casanova, A. Murgui, A. Dominguez, Neil Andrew Robert Gow, J. P. Martinez

Research output: Contribution to journalArticle

18 Citations (Scopus)

Abstract

Immunoscreening of a Candida albicans expression library resulted in the isolation of a novel gene encoding a 32.9-kDa polypeptide (288 amino acids), with 27.7% homology to the product of Saccharomyces cerevisiae YGR106c, a putative vacuolar protein. Heterozygous mutants in this gene displayed an altered budding growth pattern, characterized by the formation of chains of buds, decreasingly in size towards the apex, without separation of the daughter buds. Consequently, this gene was designated ABG1. A conditional mutant for ABG1 with the remaining allele under the control of the MET3 promoter did not grow in the presence of methionine and cysteine, demonstrating that ABG1 was essential for viability. Western analysis revealed the presence of a major 32.9-kDa band, mainly in a particulate fraction (P40) enriched in vacuoles, and tagging with green fluorescent protein confirmed that Abg1p localized to the vacuole. Vacuole inheritance has been linked to the regulation of branching frequency in C. albicans. Under repressing conditions, the conditional mutant had an increased frequency of branching under hyphal inducing conditions and an altered sensitivity to substances that interfered with cell wall assembly. Repression of ABG1 in the conditional mutant strain caused disturbance of normal size and number of vacuoles both in yeast and mycelial cells and also in the asymmetric vacuole inheritance associated with the characteristic pattern of germ tubes and branching in C. albicans. These observations indicate that ABG1 plays a key role in vacuole biogenesis, cytokinesis, and hyphal branching.

Original languageEnglish
Pages (from-to)1088-1101
Number of pages13
JournalEukaryotic Cell
Volume4
DOIs
Publication statusPublished - 2005

Keywords

  • GERM TUBE FORMATION
  • CELL-WALL
  • SACCHAROMYCES-CEREVISIAE
  • YEAST VACUOLE
  • BIOGENESIS
  • MUTANTS
  • VIRULENCE
  • EXPRESSION
  • AUTOPHAGY
  • GROWTH

Cite this

Characterization of ABG1, a novel and essential gene of Candida albicans coding for a vacuolar protein involved in cytokinesis an dhyphal branching. / Veses, V.; Casanova, M.; Murgui, A.; Dominguez, A.; Gow, Neil Andrew Robert; Martinez, J. P.

In: Eukaryotic Cell, Vol. 4, 2005, p. 1088-1101.

Research output: Contribution to journalArticle

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AU - Dominguez, A.

AU - Gow, Neil Andrew Robert

AU - Martinez, J. P.

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AB - Immunoscreening of a Candida albicans expression library resulted in the isolation of a novel gene encoding a 32.9-kDa polypeptide (288 amino acids), with 27.7% homology to the product of Saccharomyces cerevisiae YGR106c, a putative vacuolar protein. Heterozygous mutants in this gene displayed an altered budding growth pattern, characterized by the formation of chains of buds, decreasingly in size towards the apex, without separation of the daughter buds. Consequently, this gene was designated ABG1. A conditional mutant for ABG1 with the remaining allele under the control of the MET3 promoter did not grow in the presence of methionine and cysteine, demonstrating that ABG1 was essential for viability. Western analysis revealed the presence of a major 32.9-kDa band, mainly in a particulate fraction (P40) enriched in vacuoles, and tagging with green fluorescent protein confirmed that Abg1p localized to the vacuole. Vacuole inheritance has been linked to the regulation of branching frequency in C. albicans. Under repressing conditions, the conditional mutant had an increased frequency of branching under hyphal inducing conditions and an altered sensitivity to substances that interfered with cell wall assembly. Repression of ABG1 in the conditional mutant strain caused disturbance of normal size and number of vacuoles both in yeast and mycelial cells and also in the asymmetric vacuole inheritance associated with the characteristic pattern of germ tubes and branching in C. albicans. These observations indicate that ABG1 plays a key role in vacuole biogenesis, cytokinesis, and hyphal branching.

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KW - CELL-WALL

KW - SACCHAROMYCES-CEREVISIAE

KW - YEAST VACUOLE

KW - BIOGENESIS

KW - MUTANTS

KW - VIRULENCE

KW - EXPRESSION

KW - AUTOPHAGY

KW - GROWTH

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