AP-2-dependent endocytic recycling of the chitin synthase Chs3 regulates polarized growth in Candida albicans

H. C. Knafler, I. I. Smaczynska-de Rooij, L. A. Walker, K. K. Lee, N. A.R. Gow, K. R. Ayscougha

Research output: Contribution to journalArticle

Abstract

The human fungal pathogen Candida albicans is known to require endocytosis to enable its adaptation to diverse niches and to maintain its highly polarized hyphal growth phase. While studies have identified changes in transcription leading to the synthesis and secretion of new proteins to facilitate hyphal growth, effective maintenance of hyphae also requires concomitant removal or relocalization of other cell surface molecules. The key molecules which must be removed from the cell surface, and the mechanisms behind this, have, however, remained elusive. In this study, we show that the AP-2 endocytic adaptor complex is required for the internalization of the major cell wall biosynthesis enzyme Chs3. We demonstrate that this interaction is mediated by the AP-2 mu subunit (Apm4) YXXΦ binding domain. We also show that in the absence of Chs3 recycling via AP-2, cells have abnormal cell wall composition, defective polarized cell wall deposition, and morphological defects. The study also highlights key distinctions between endocytic requirements of growth at yeast buds compared to that at hyphal tips and different requirements of AP-2 in maintaining the polarity of mannosylated proteins and ergosterol at hyphal tips. Together, our findings highlight the importance of correct cell wall deposition in cell shape maintenance and polarized growth and the key regulatory role of endocytic recycling via the AP-2 complex. IMPORTANCE Candida albicans is a human commensal yeast that can cause significant morbidity and mortality in immunocompromised individuals. Within humans, C. albicans can adopt different morphologies as yeast or filamentous hyphae and can occupy different niches with distinct temperatures, pHs, CO2 levels, and nutrient availability. Both morphological switching and growth in different environments require cell surface remodelling, which involves both the addition of newly synthesized proteins as well as the removal of other proteins. In our study, we demonstrate the importance of an adaptor complex AP-2 in internalizing and recycling a specific cell surface enzyme to maintain effective polarized hyphal growth. Defects in formation of the complex or in its ability to interact directly with cargo inhibit enzyme uptake and lead to defective cell walls and aberrant hyphal morphology. Our data indicate that the AP-2 adaptor plays a central role in regulating cell surface composition in Candida.

Original languageEnglish
Article numbere02421-18
JournalmBio
Volume10
Issue number2
Early online date19 Mar 2019
DOIs
Publication statusE-pub ahead of print - 19 Mar 2019

Fingerprint

Chitin Synthase
Candida albicans
Cell Wall
Growth
Hyphae
Yeasts
Recycling
Proteins
Enzymes
Maintenance
Ergosterol
Cell Shape
Endocytosis
Candida
Morbidity
Food
Temperature
Mortality

Keywords

  • Candida albicans
  • Cell polarity
  • Cell wall
  • Endocytosis
  • Membrane trafficking
  • Yeasts
  • Chitin Synthase/metabolism
  • Candida albicans/enzymology
  • Hyphae/enzymology
  • Adaptor Protein Complex 2/metabolism
  • CONGO RED
  • yeasts
  • PLASMA-MEMBRANE
  • CELL-WALL
  • endocytosis
  • cell polarity
  • STRUCTURAL EXPLANATION
  • membrane trafficking
  • CALCOFLUOR WHITE
  • MEDIATED ENDOCYTOSIS
  • GENE DELETION
  • cell wall
  • SORTING SIGNALS
  • CLATHRIN
  • HYPHAL GROWTH

ASJC Scopus subject areas

  • Virology
  • Microbiology

Cite this

Knafler, H. C., Smaczynska-de Rooij, I. I., Walker, L. A., Lee, K. K., Gow, N. A. R., & Ayscougha, K. R. (2019). AP-2-dependent endocytic recycling of the chitin synthase Chs3 regulates polarized growth in Candida albicans. mBio, 10(2), [e02421-18]. https://doi.org/10.1128/mBio.02421-18

AP-2-dependent endocytic recycling of the chitin synthase Chs3 regulates polarized growth in Candida albicans. / Knafler, H. C.; Smaczynska-de Rooij, I. I.; Walker, L. A.; Lee, K. K.; Gow, N. A.R.; Ayscougha, K. R.

In: mBio, Vol. 10, No. 2, e02421-18, 19.03.2019.

Research output: Contribution to journalArticle

Knafler, HC, Smaczynska-de Rooij, II, Walker, LA, Lee, KK, Gow, NAR & Ayscougha, KR 2019, 'AP-2-dependent endocytic recycling of the chitin synthase Chs3 regulates polarized growth in Candida albicans' mBio, vol. 10, no. 2, e02421-18. https://doi.org/10.1128/mBio.02421-18
Knafler, H. C. ; Smaczynska-de Rooij, I. I. ; Walker, L. A. ; Lee, K. K. ; Gow, N. A.R. ; Ayscougha, K. R. / AP-2-dependent endocytic recycling of the chitin synthase Chs3 regulates polarized growth in Candida albicans. In: mBio. 2019 ; Vol. 10, No. 2.
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abstract = "The human fungal pathogen Candida albicans is known to require endocytosis to enable its adaptation to diverse niches and to maintain its highly polarized hyphal growth phase. While studies have identified changes in transcription leading to the synthesis and secretion of new proteins to facilitate hyphal growth, effective maintenance of hyphae also requires concomitant removal or relocalization of other cell surface molecules. The key molecules which must be removed from the cell surface, and the mechanisms behind this, have, however, remained elusive. In this study, we show that the AP-2 endocytic adaptor complex is required for the internalization of the major cell wall biosynthesis enzyme Chs3. We demonstrate that this interaction is mediated by the AP-2 mu subunit (Apm4) YXXΦ binding domain. We also show that in the absence of Chs3 recycling via AP-2, cells have abnormal cell wall composition, defective polarized cell wall deposition, and morphological defects. The study also highlights key distinctions between endocytic requirements of growth at yeast buds compared to that at hyphal tips and different requirements of AP-2 in maintaining the polarity of mannosylated proteins and ergosterol at hyphal tips. Together, our findings highlight the importance of correct cell wall deposition in cell shape maintenance and polarized growth and the key regulatory role of endocytic recycling via the AP-2 complex. IMPORTANCE Candida albicans is a human commensal yeast that can cause significant morbidity and mortality in immunocompromised individuals. Within humans, C. albicans can adopt different morphologies as yeast or filamentous hyphae and can occupy different niches with distinct temperatures, pHs, CO2 levels, and nutrient availability. Both morphological switching and growth in different environments require cell surface remodelling, which involves both the addition of newly synthesized proteins as well as the removal of other proteins. In our study, we demonstrate the importance of an adaptor complex AP-2 in internalizing and recycling a specific cell surface enzyme to maintain effective polarized hyphal growth. Defects in formation of the complex or in its ability to interact directly with cargo inhibit enzyme uptake and lead to defective cell walls and aberrant hyphal morphology. Our data indicate that the AP-2 adaptor plays a central role in regulating cell surface composition in Candida.",
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note = "We thank Andrew Peden, Simon Johnston, and Josh Parker for critical reading of the manuscript. We also thank Gillian Milne at the Microscopy and Histology Core Facility at the University of Aberdeen for expert assistance with TEM and Craig Murdoch at the School of Clinical Dentistry, University of Sheffield, for training and support in carrying out the biofilm assays. Imaging was undertaken in part at the Wolfson Light Microscopy Facility at the University of Sheffield, supported by grant number MR/K015753/1. H.C.K. is funded by the BBSRC White Rose DTP to the Universities of Sheffield, Leeds and York (BB/M011151/1); I.I.S.-D.R. is funded by BBSRC grant BB/N007581/1. N.A.R.G. was supported by The Wellcome Trust (101873, 200208, 097377), and L.A.W. and K.K.L. were supported by the MRC Centre for Medical Mycology (MR/N006364/1).",
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T1 - AP-2-dependent endocytic recycling of the chitin synthase Chs3 regulates polarized growth in Candida albicans

AU - Knafler, H. C.

AU - Smaczynska-de Rooij, I. I.

AU - Walker, L. A.

AU - Lee, K. K.

AU - Gow, N. A.R.

AU - Ayscougha, K. R.

N1 - We thank Andrew Peden, Simon Johnston, and Josh Parker for critical reading of the manuscript. We also thank Gillian Milne at the Microscopy and Histology Core Facility at the University of Aberdeen for expert assistance with TEM and Craig Murdoch at the School of Clinical Dentistry, University of Sheffield, for training and support in carrying out the biofilm assays. Imaging was undertaken in part at the Wolfson Light Microscopy Facility at the University of Sheffield, supported by grant number MR/K015753/1. H.C.K. is funded by the BBSRC White Rose DTP to the Universities of Sheffield, Leeds and York (BB/M011151/1); I.I.S.-D.R. is funded by BBSRC grant BB/N007581/1. N.A.R.G. was supported by The Wellcome Trust (101873, 200208, 097377), and L.A.W. and K.K.L. were supported by the MRC Centre for Medical Mycology (MR/N006364/1).

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N2 - The human fungal pathogen Candida albicans is known to require endocytosis to enable its adaptation to diverse niches and to maintain its highly polarized hyphal growth phase. While studies have identified changes in transcription leading to the synthesis and secretion of new proteins to facilitate hyphal growth, effective maintenance of hyphae also requires concomitant removal or relocalization of other cell surface molecules. The key molecules which must be removed from the cell surface, and the mechanisms behind this, have, however, remained elusive. In this study, we show that the AP-2 endocytic adaptor complex is required for the internalization of the major cell wall biosynthesis enzyme Chs3. We demonstrate that this interaction is mediated by the AP-2 mu subunit (Apm4) YXXΦ binding domain. We also show that in the absence of Chs3 recycling via AP-2, cells have abnormal cell wall composition, defective polarized cell wall deposition, and morphological defects. The study also highlights key distinctions between endocytic requirements of growth at yeast buds compared to that at hyphal tips and different requirements of AP-2 in maintaining the polarity of mannosylated proteins and ergosterol at hyphal tips. Together, our findings highlight the importance of correct cell wall deposition in cell shape maintenance and polarized growth and the key regulatory role of endocytic recycling via the AP-2 complex. IMPORTANCE Candida albicans is a human commensal yeast that can cause significant morbidity and mortality in immunocompromised individuals. Within humans, C. albicans can adopt different morphologies as yeast or filamentous hyphae and can occupy different niches with distinct temperatures, pHs, CO2 levels, and nutrient availability. Both morphological switching and growth in different environments require cell surface remodelling, which involves both the addition of newly synthesized proteins as well as the removal of other proteins. In our study, we demonstrate the importance of an adaptor complex AP-2 in internalizing and recycling a specific cell surface enzyme to maintain effective polarized hyphal growth. Defects in formation of the complex or in its ability to interact directly with cargo inhibit enzyme uptake and lead to defective cell walls and aberrant hyphal morphology. Our data indicate that the AP-2 adaptor plays a central role in regulating cell surface composition in Candida.

AB - The human fungal pathogen Candida albicans is known to require endocytosis to enable its adaptation to diverse niches and to maintain its highly polarized hyphal growth phase. While studies have identified changes in transcription leading to the synthesis and secretion of new proteins to facilitate hyphal growth, effective maintenance of hyphae also requires concomitant removal or relocalization of other cell surface molecules. The key molecules which must be removed from the cell surface, and the mechanisms behind this, have, however, remained elusive. In this study, we show that the AP-2 endocytic adaptor complex is required for the internalization of the major cell wall biosynthesis enzyme Chs3. We demonstrate that this interaction is mediated by the AP-2 mu subunit (Apm4) YXXΦ binding domain. We also show that in the absence of Chs3 recycling via AP-2, cells have abnormal cell wall composition, defective polarized cell wall deposition, and morphological defects. The study also highlights key distinctions between endocytic requirements of growth at yeast buds compared to that at hyphal tips and different requirements of AP-2 in maintaining the polarity of mannosylated proteins and ergosterol at hyphal tips. Together, our findings highlight the importance of correct cell wall deposition in cell shape maintenance and polarized growth and the key regulatory role of endocytic recycling via the AP-2 complex. IMPORTANCE Candida albicans is a human commensal yeast that can cause significant morbidity and mortality in immunocompromised individuals. Within humans, C. albicans can adopt different morphologies as yeast or filamentous hyphae and can occupy different niches with distinct temperatures, pHs, CO2 levels, and nutrient availability. Both morphological switching and growth in different environments require cell surface remodelling, which involves both the addition of newly synthesized proteins as well as the removal of other proteins. In our study, we demonstrate the importance of an adaptor complex AP-2 in internalizing and recycling a specific cell surface enzyme to maintain effective polarized hyphal growth. Defects in formation of the complex or in its ability to interact directly with cargo inhibit enzyme uptake and lead to defective cell walls and aberrant hyphal morphology. Our data indicate that the AP-2 adaptor plays a central role in regulating cell surface composition in Candida.

KW - Candida albicans

KW - Cell polarity

KW - Cell wall

KW - Endocytosis

KW - Membrane trafficking

KW - Yeasts

KW - Chitin Synthase/metabolism

KW - Candida albicans/enzymology

KW - Hyphae/enzymology

KW - Adaptor Protein Complex 2/metabolism

KW - CONGO RED

KW - yeasts

KW - PLASMA-MEMBRANE

KW - CELL-WALL

KW - endocytosis

KW - cell polarity

KW - STRUCTURAL EXPLANATION

KW - membrane trafficking

KW - CALCOFLUOR WHITE

KW - MEDIATED ENDOCYTOSIS

KW - GENE DELETION

KW - cell wall

KW - SORTING SIGNALS

KW - CLATHRIN

KW - HYPHAL GROWTH

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