School of Medical Sciences, University of Aberdeen, Institute of Medical Sciences, Foresterhill, Aberdeen AB25 2ZD, UK
Correspondence
Neil A. R. Gow
n.gow{at}abdn.ac.uk
![]() |
ABSTRACT |
---|
![]() ![]() ![]() ![]() ![]() ![]() ![]() |
---|
The GenBank/EMBL/DDBJ accession number for the sequence reported in this paper is AY569336.
Present address: Molecular Genetics and Oncology Group, Department of Clinical Dental Sciences, University of Liverpool, Edwards Building, Daulby Street, Liverpool L69 3GN, UK.
![]() |
INTRODUCTION |
---|
![]() ![]() ![]() ![]() ![]() ![]() ![]() |
---|
In C. albicans, CaChs3p expression is increased in hyphae, which have more chitin than yeast cells (Munro et al., 1998). CaChs1p synthesizes the chitin of the primary septum, is expressed at a low level in both yeast and hyphal cells, and has a potential role in lateral cell-wall stability (Munro et al., 1998
, 2001
, 2003
). CaChs2p is a non-essential enzyme and its expression is increased during the yeast-to-hyphal transition (Gow et al., 1994
). CaChs8p also contributes to in vitro chitin synthase activity in yeast and hyphal cells and has highest identity to CaChs2p (Munro et al., 2003
). Therefore, chitin synthesis is regulated temporally and spatially at the transcriptional and post-translational levels (Roncero, 2002
; Munro & Gow, 2001
).
The spatial and temporal regulation of Chs3p has been studied in S. cerevisiae and involves a number of ancillary proteins including ScChs4p, ScChs5p, ScChs6p and ScChs7p (Trilla et al., 1997, 1999
; Santos et al., 1997
; Ziman et al., 1998
), septins and Bni4p (DeMarini et al., 1997
). In both C. albicans and S. cerevisiae, Chs4p is involved directly or indirectly in the activation of Chs3p (Sudoh et al., 1999
; Ono et al., 2000
). ScChs5p is required for targeting of ScChs3p to polarized growth sites and for cell fusion during mating (Santos et al., 1997
; Santos & Snyder, 2003
). ScChs6p is involved in the spatial regulation and recycling of ScChs3p (Ziman et al., 1998
), while ScChs7p is required for the export of ScChs3p from the endoplasmic reticulum to the plasma membrane (Trilla et al., 1999
). ScChs3p is transported through the secretory pathway to the plasma membrane where chitin synthesis takes place, and can also be internalized again via ligand-induced endocytosis (Ziman et al., 1996
). ScChs3p can also be mobilized from internal stores in response to cellular stress (Valdivia & Schekman, 2003
).
In S. cerevisiae, chitin deposition at the bud site is linked to the septin neck filaments, a family of proteins that have roles in septation and cytokinesis. The septin proteins localize to the incipient bud site and are ordered into higher structures to form a ring of neck filaments. The actin-myosin ring contracts and then the two neck filaments split, resulting in cytokinesis so that the mother and daughter cells each inherit a single septin ring (Longtine et al., 1996; Lippincott et al., 2001
; Kinoshita, 2003
). In S. cerevisiae there are seven septin genes: CDC3, CDC10, CDC11, CDC12, SHS1/SEP7, SPR3 and SPR28. The latter two genes are expressed only in the sporulation phase. In C. albicans, homologues of all seven septin genes have been recognized. The CaCDC3 and CaCDC10 genes are capable of complementing defects in the respective S. cerevisiae genes and are thus believed to perform similar roles (Di Domenico et al., 1994
). Similar to the S. cerevisiae septins, CaCDC3 and CaCDC12 are essential for viability; however, the Cacdc10
and Cacdc11
mutants were viable but displayed conditional defects in cytokinesis, chitin localization and bud morphology (Warenda & Konopka, 2002
). The C. albicans Cdc3p and Cdc11p homologues have been localized during the growth of yeast, pseudohyphae and hyphae (Sudbery, 2001
). In true hyphae, an early and transient septin ring is formed that lacks Cdc3p, which does not participate in cytokinesis later in the cell cycle (Sudbery, 2001
). Cacdc11
mutants were defective in ability to form hyphae on solid agar (Warenda & Konopka, 2002
).
In S. cerevisiae, the septin Cdc10p interacts with Chs4p via Bni4p (bud neck involved). The ScBni4p protein is required for the correct targeting of ScChs3p and its activator ScChs4p to the bud-neck region. Mutants deleted in ScBNI4 had delocalized chitin deposition, with an aberrant cell morphology and enlarged bud necks (DeMarini et al., 1997). Immunolocalization experiments confirmed that ScBni4p was normally localized to the mother-bud-neck region and that localization was dependent upon the presence of septins but not ScChs4p or ScChs3p (DeMarini et al., 1997
).
Yeast dihybrid analysis demonstrated that ScBni4p interacts with ScGlc7p (Uetz et al., 2000). ScGlc7p is a protein phosphatase type 1 (PP1) involved in a range of physiological activities including cell polarity, cell-wall integrity and morphology (Andrews & Stark, 2000
). ScBni4p is required for the localization of ScGlc7p to the incipient bud neck but not for targeting to the neck at cytokinesis (Kozubowski et al., 2003
). Although the precise role for this phosphatase at the bud neck is not known, it could facilitate the association of ScBni4p with the septins. ScBni4p is also believed to be necessary for the targeting of ScCrh2p, a glycosylphosphatidylinositol (GPI)-anchored mannoprotein involved in cell-wall assembly, at cytokinesis (Rodriguez-Pena et al., 2002
).
Chitin synthesis in C. albicans is significantly different from that in S. cerevisiae in a number of respects (Munro & Gow, 2001; Munro et al., 2003
), and we therefore examined the role of CaBni4p in growth and cellular morphogenesis in C. albicans. Here we describe the isolation and analysis of the C. albicans BNI4 homologue by reverse genetics. Disruption of CaBNI4 resulted in cells that had a phenotype that was again different, in some aspects, to that of the bni4 mutation in S. cerevisiae. These were depleted in total chitin and had abnormal cell shape and bud-scar morphology and defects in hypha formation when growing on solid surfaces.
![]() |
METHODS |
---|
![]() ![]() ![]() ![]() ![]() ![]() ![]() |
---|
|
Bacterial strains and plasmids.
Escherichia coli DH5 and XL1Blue (Stratagene) were used for preparation of plasmid DNA. Plasmids pGEM-T, pBluescript and pBR322 (Promega) were used for cloning gene fragments. For sequencing and disruption cassette construction, the CaBNI4 gene sequence was cloned as two overlapping fragments. Attempts to clone the whole gene were unsuccessful due to frequent recombination events in E. coli.
Plasmids p5921 and pMB7 were used for construction of Ura-blaster cassettes (Fonzi & Irwin, 1993; Gow et al., 1994
). A 543 bp fragment from the middle of the coding region of CaBNI4 and a 543 bp fragment covering the 3' coding and non-coding region of CaBNI4 were obtained by PCR using restriction-site-ended primers, to facilitate the directed cloning into plasmids p5921 and pMB7 following digestion with the corresponding restriction endonucleases (Table 2
). The resulting disruption plasmids, named p59BN1 and pMBN2, were derived from p5921 and pMB7 respectively, and were used to transform C. albicans strain CAI-4. Homologous integration resulted in the disruption of nucleotides 36304833 from the 4968 nucleotide ORF (corresponding to amino acids 11201611 out of 1655 total). This region corresponded to the putative and conserved RVXF Glc7-binding motif that associates with the hydrophobic groove of the catalytic subunit, PP1c (Egloff et al., 1997
), that is believed to be essential for Bni4p function.
|
Analysis of nucleic acids.
C. albicans genomic DNA was extracted using the mechanical lysis method by vortexing in Triton mix [2 % (w/v) Triton X-100, 1 % (w/v) SDS, 100 mM NaCl, 10 mM Tris/HCl, pH 8·0 and 1 mM EDTA] with glass beads (Sigma, 425600 µm in diameter) followed by phenol/chloroform extraction. DNA samples were digested with restriction endonucleases, separated on a 0·81 % (w/v) agarose gel and transferred to nylon membranes. Total RNA was extracted from 50100 mg (wet wt) yeast and hyphal C. albicans cells using the Qiagen RNeasy mini extraction kit following lysis in a Hybaid Ribolyser. RNA samples were heated at 50 °C for 60 min in 55 % DMSO, 1·1 M Glyoxal, separated on a 1·4 % (w/v) non-denaturing agarose gel and again transferred to nylon membranes.
Southern and Northern blots were probed with -32P-radiolabelled fragments of CaBNI4. DNA sequencing was performed using the dideoxy-termination method (Sanger et al., 1977
) and sequences were aligned and analysed using the GCG v.10 software from HGMP (http://www.hgmp.mrc.ac.uk). Northern blots were re-probed with a radiolabelled CaTEF3 fragment as loading controls (Di Domenico et al., 1992
). The entire gene was sequenced and submitted to GenBank: accession number AY569336.
Chitin contents.
C. albicans cell-wall chitin was determined by the enzymic method (Bulawa et al., 1986; Mellado et al., 1996
; Munro et al., 1998
) and acid hydrolysis methods described previously (Kapteyn et al., 2000
). Triplicate samples of 50100 mg (wet wt) yeast cells were used in the enzymic analysis and 2030 mg (wet wt) cell-wall material were used for the acid-hydrolysis method.
Morphological observations.
Yeast and hyphal cells were fixed in 10 % (v/v) neutral buffered formalin (Sigma). Chitin was stained with 50 µg Calcofluor ml1 (Sigma, fluorescent brightener 28). Transmission electron microscopy (TEM) was performed on exponential-phase yeast cells after fixation with 3 % glutaraldehyde (v/v) in 0·1 M sodium phosphate buffer pH 7·4. Secondary fixation was performed in 1 % osmium tetroxide in distilled H2O and cells were embedded in TAAB resin. Ultrathin silver or gold defracting sections were stained with uranyl acetate and lead citrate and examined with a Philips 301 transmission electron microscope at an accelerating voltage of 80 kV.
![]() |
RESULTS |
---|
![]() ![]() ![]() ![]() ![]() ![]() ![]() |
---|
|
|
|
|
|
|
|
![]() |
DISCUSSION |
---|
![]() ![]() ![]() ![]() ![]() ![]() ![]() |
---|
In S. cerevisiae it has been shown that Bni4p interacts with Chs4p and the septin Cdc10p whilst Chs4p interacts with Cdc12p (DeMarini et al., 1997). Therefore, if the same is true in C. albicans, then the Chs4pCdc12p interaction may be sufficient for targeting of Chs3p to the bud site since the bud neck and primary septum were not markedly affected in the mutant. To date no other ScBNI4 homologue has been identified in the C. albicans genome sequencing project and no homologues were identified here by low-stringency hybridization; therefore, no evidence exists for the formal possibility that function of Bni4p can be achieved by an alternative homologous protein.
Changes in the structural integrity of the Candida cell wall are expressed as changes in the sensitivity of cells to various inhibitors that stress the wall in various ways. Sensitivity to Hygromycin B, SDS, Calcofluor and NaCl have been shown to depend on the permeability, porosity or chitin content of the cell wall, and several glycosylation or chitin synthase mutants have increased sensitivity to these compounds (Timpel et al., 1998, 2000
; Warit et al., 2000
; Binley et al., 1999
; Navarro-Garcia et al., 1995
). The Cabni4
/Cabni4
mutant had decreased sensitivity to Calcofluor White, reflecting the reduced chitin content, and increased sensitivity to SDS that may reflect reduced integrity of the wall.
We have demonstrated a reduction in cell-wall chitin in the Cabni4/Cabni4
null mutant, indicating that in C. albicans, Bni4p is involved in targeting of Chs3p to the cell surface. However, disruption of CaBNI4 caused loss of chitin over the whole cell wall as visualized by Calcofluor staining. We propose that CaBni4p may have a global, septin/bud-neck-independent role in the deposition of chitin, by either the targeting or activation of the Chs3p chitin synthase enzyme.
The lemon shape of the bni4/bni4
mutants suggests that Chs3p targeting and chitin content are vital for normal yeast morphology. The C. albicans bni4
/bni4
mutant was also reduced in hyphal formation on 20 % serum agar. Germ-tube formation was, however, normal in both Lee's and 20 % serum liquid media. A number of other cell-wall mutants of C. albicans such as int1
(Gale et al., 1998
), hwp1
(Tsuchimori et al., 2000
) and cdc10
and cdc11
septin mutants (Warenda et al., 2003
) have been found to respond differently to hypha-inducing signals in liquid and solid media. For example, mutants of the non-essential septins cdc10
and cdc11
were only mildly affected for hyphal growth in liquid culture, whereas hyphal growth was profoundly affected on solid media (Warenda et al., 2003
). The Cabni4
/Cabni4
mutant described here was more severely defective in hyphal growth on solid media even in comparison with the cdc10
and cdc11
mutants. The zinc finger transcription factor Czf1 has also been shown to be involved in the regulation of hyphal formation only when the cell was in contact with solid medium (Brown et al., 1999
). These observations suggest that different or additional regulatory processes are involved in the formation of hyphal cells on solid and in liquid media and that Bni4p may be important in the deposition of chitin that maintains the filamentous cell shape when hyphal cells are growing on surfaces.
It is not known what proteins are involved in the targeting of Chs enzymes, other than Chs3p, to specific cellular locations. Analysis of disrupted CHS genes in C. albicans suggests that Chs3p, Chs2p and Chs1p may all participate in the formation of chitin in the cell wall of hyphae (Gow et al., 1994; Bulawa et al., 1995
; Munro et al., 2001
). The activity of these enzymes is not thought to be dependent on Chs4p. No association of Bni4p or Chs4p with other Chs proteins was identified by dihybrid analysis in S. cerevisiae (DeMarini et al., 1997
). However, the possibility that Bni4p is involved in the targeting of other chitin synthase enzymes to the surface has yet to be fully explored. Although the phenotype of the bni4
/bni4
mutant of C. albicans differs significantly in some respects from the equivalent S. cerevisiae mutant, our data support the evidence that disruption of the chitin synthase cell-targeting machinery influences the composition, architecture and function of the fungal cell wall.
![]() |
ACKNOWLEDGEMENTS |
---|
![]() |
REFERENCES |
---|
![]() ![]() ![]() ![]() ![]() ![]() ![]() |
---|
Binley, K. M., Radcliffe, P. A., Trevethick, J., Duffy, K. A. & Sudbery, P. E. (1999). The yeast PRS3 gene is required for cell integrity, cell cycle arrest upon nutrient deprivation, ion homeostasis and the proper organization of the actin cytoskeleton. Yeast 14, 14591469.[CrossRef]
Brown, D. H., Jr, Giusani, A. D., Chen, X. & Kumamoto, C. A. (1999). Filamentous growth of Candida albicans in response to physical environmental cues and its regulation by the unique CZF1 gene. Mol Microbiol 34, 651662.[CrossRef][Medline]
Bulawa, C. E., Slater, M., Cabib, E., Au-Young, J., Sburlati, A., Adair, W. L. & Robbins, P. W. (1986). The S. cerevisiae structural gene for chitin synthase is not required for chitin synthesis in vivo. Cell 46, 213225.[Medline]
Bulawa, C. E., Miller, D. W., Henry, L. K. & Becker, J. M. (1995). Attenuated virulence of chitin-deficient mutants of Candida albicans. Proc Natl Acad Sci U S A 92, 1057010574.[Abstract]
Cassola, A., Parrot, M., Silberstein, S., Magee, B. B., Passerson, S., Giasson, L. & Cantore, M. L. (2004). Candida albicans lacking the gene encoding the regulatory subunit of protein kinase A displays a defect in hyphal formation and an altered localization of the catalytic subunit. Eukaryot Cell 3, 190199.
DeMarini, D. J., Adams, A. E. M., Fares, H., De Virgilio, C., Valle, G., Chuang, J. S. & Pringle, J. R. (1997). A septin-based hierarchy of proteins required for localized deposition of chitin in the Saccharomyces cerevisiae cell wall. J Cell Biol 139, 7593.
Di Domenico, B. J., Lupisella, J., Sandbaken, M. & Chakraburtty, K. (1992). Isolation and sequence analysis of the gene encoding translation elongation factor 3 from Candida albicans. Yeast 8, 337352.[Medline]
Di Domenico, B. J., Brown, N. H., Lupisella, J., Greene, J. R., Yanko, M. & Koltin, Y. (1994). Homologues of the yeast neck filament associated genes: isolation and sequence analysis of Candida albicans CDC3 and CDC10. Mol Gen Genet 242, 689698.[Medline]
Egloff, M. P., Johnson, D. F., Moorhead, G., Cohen, P. T., Cohen, P. & Barford, D. (1997). Structural basis for the recognition of regulatory subunits by the catalytic subunit of protein phosphatase 1. EMBO J 16, 18761887.
Fonzi, W. A. & Irwin, M. Y. (1993). Isogenic strain construction and gene mapping in Candida albicans. Genetics 134, 717728.
Gale, C. A., Bendel, C. M., McClellan, M., Hauser, M., Becker, J. M., Berman, J. & Hostetter, M. K. (1998). Linkage of adhesion, filamentous growth, and virulence in Candida albicans to a single gene, INT1. Science 279, 13551358.
Gow, N. A. R., Robbins, P. W., Lester, J. W., Brown, A. J., Fonzi, W. A., Chapman, T. & Kinsman, O. S. (1994). A hyphal-specific chitin synthase gene (CHS2) is not essential for growth, dimorphism, or virulence of Candida albicans. Proc Natl Acad Sci U S A 91, 62166220.[Abstract]
Kapteyn, J. C., Hoyer, L. L., Hecht, J. E. & 6 other authors (2000). The cell wall architecture of Candida albicans wild-type cells and cell wall-defective mutants. Mol Microbiol 35, 601611.[CrossRef][Medline]
Kinoshita, M. (2003). The septins. Genome Biol 4, 236 (doi:10.1186/gb-2003-4-11-236).[CrossRef][Medline]
Kozubowski, L., Panek, H., Rosenthal, A., Bloecher, A., DeMarini, D. J. & Tatchell, K. (2003). A Bni4-Glc7 phosphatase complex that recruits chitin synthase to the site of bud emergence. Mol Biol Cell 14, 2639.
Lee, K. L., Buckley, H. R. & Campbell, H. R. (1975). An amino acid liquid synthetic medium for development of mycelial and yeast forms of C. albicans. Sabouraudia 13, 148153.[Medline]
Lippincott, J., Shannon, K. B., Shou, W., Deshaies, R. J. & Li, R. (2001). The TEM1 small GTPase controls actomyosin and septin dynamics during cytokinesis. J Cell Sci 114, 13791386.
Longtine, M. S., DeMarini, D. J., Valencik, M. L., Al-Awar, O. S., Fares, H., De Virgilio, C. & Pringle, J. R. (1996). The septins: roles in cytokinesis and other processes. Curr Opin Cell Biol 8, 106119.[CrossRef][Medline]
Mellado, E., Specht, C. A., Robbins, P. W. & Holden, D. W. (1996). Cloning and characterization of chsD, a chitin synthase-like gene of Aspergillus fumigatus. FEMS Lett 143, 6976.
Munro, C. A. & Gow, N. A. R. (2001). Chitin synthesis in human pathogenic fungi. Med Mycol 39S, 4154.
Munro, C. A., Schofield, D. A., Gooday, G. W. & Gow, N. A. R. (1998). Regulation of chitin synthesis during dimorphic growth of Candida albicans. Microbiology 144, 391401.[Medline]
Munro, C. A., Winter, K., Buchan, A., Henry, K., Becker, J. M., Brown, A. J., Bulawa, C. E. & Gow, N. A. R. (2001). Chs1 of Candida albicans is an essential chitin synthase required for synthesis of the septum and for cell integrity. Mol Microbiol 39, 14141426.[CrossRef][Medline]
Munro, C. A., Whitton, R. K., Hughes, H. B., Rella, M., Selvaggini, S. & Gow, N. A. R. (2003). CHS8-a fourth chitin synthase gene of Candida albicans contributes to in vitro chitin synthase activity, but is dispensable for growth. Fungal Gen Biol 40, 146158.[CrossRef]
Navarro-Garcia, F., Sanchez, M., Pla, J. & Nombela, C. (1995). Functional characterization of the MKC1 gene of Candida albicans, which encodes a mitogen-activated protein kinase homolog related to cell integrity. Mol Cell Biol 15, 21972206.[Abstract]
Ono, N., Yabe, T., Sudoh, M., Nakajima, T., Yamada-Okabe, T., Arisawa, M. & Yamada-Okabe, H. (2000). The yeast Chs4 protein stimulates the trypsin-sensitive activity of chitin synthase 3 through an apparent protein-protein interaction. Microbiology 146, 385391.[Medline]
Rodriguez-Pena, J. M., Rodriguez, C., Alvarez, A., Nombela, C. & Arroyo, J. (2002). Mechanisms for targeting of the Saccharomyces cerevisiae GPI-anchored cell wall protein Crh2p to polarised growth sites. J Cell Sci 115, 25492558.
Roncero, C. (2002). The genetic complexity of chitin synthesis in fungi. Curr Genet 41, 367378.[CrossRef][Medline]
Sanger, F., Nicklen, S. & Coulson, A. R. (1977). DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci U S A 74, 54635467.[Abstract]
Sanglard, D., Hube, B., Monod, M., Odds, F. C. & Gow, N. A. R. (1997). A triple deletion of the secreted aspartyl proteinase genes SAP4, SAP5, and SAP6 of Candida albicans causes attenuated virulence. Infect Immun 65, 35393546.[Abstract]
Santos, B. & Snyder, M. (2003). Specific protein targeting during cell differentiation: polarized localization of Fus1p during mating depends on Chs5p in Saccharomyces cerevisiae. Eukaryot Cell 2, 821825.
Santos, B., Duran, A. & Valdivieso, M. H. (1997). CHS5, a gene involved in chitin synthesis and mating in Saccharomyces cerevisiae. Mol Cell Biol 17, 24852496.[Abstract]
Sudbery, P. E. (2001). The germ tubes of Candida albicans hyphae and pseudohyphae show different patterns of septin ring localization. Mol Microbiol 41, 1931.[CrossRef][Medline]
Sudoh, M., Tatsuno, K., Ono, N., Ohta, A., Chibana, H., Yamada-Okabe, H. & Arisawa, M. (1999). The Candida albicans CHS4 gene complements a Saccharomyces cerevisiae skt5/chs4 mutation and is involved in chitin biosynthesis. Microbiology 145, 16131622.[Medline]
Timpel, C., Strahl-Bolsinger, S., Ziegelbauer, K. & Ernst, J. F. (1998). Multiple functions of Pmt1p-mediated protein O-mannosylation in the fungal pathogen Candida albicans. J Biol Chem 273, 2083720846.
Timpel, C., Zink, S., Strahl-Bolsinger, S., Schroppel, K. & Ernst, J. (2000). Morphogenesis, adhesive properties, and antifungal resistance depend on the Pmt6 protein mannosyltransferase in the fungal pathogen Candida albicans. J Bacteriol 182, 30633071.
Trilla, J. A., Cos, T., Duran, A. & Roncero, C. (1997). Characterization of CHS4 (CAL2), a gene of Saccharomyces cerevisiae involved in chitin biosynthesis and allelic to SKT5 and CSD4. Yeast 13, 795807.[CrossRef][Medline]
Trilla, J. A., Duran, A. & Roncero, C. (1999). Chs7p, a new protein involved in the control of protein export from the endoplasmic reticulum that is specifically engaged in the regulation of chitin synthesis in Saccharomyces cerevisiae. J Cell Biol 145, 11531163.
Tsuchimori, N., Sharkey, L. L., Fonzi, W. A., French, S. W., Edwards, J. E., Jr & Filler, S. G. (2000). Reduced virulence of HWP1-deficient mutants of Candida albicans and their interactions with host cells. Infect Immun 68, 19972002.
Uetz, P., Giot, L., Cagney, G. & 16 other authors (2000). A comprehensive analysis of protein-protein interactions in Saccharomyces cerevisiae. Nature 403, 623627.[CrossRef][Medline]
Valdivia, R. H. & Schekman, R. (2003). The yeasts Rho1p and Pkc1p regulate the transport of chitin synthase III (Chs3p) from internal stores to the plasma membrane. Proc Natl Acad Sci U S A 100, 1028710292.
Warenda, A. J. & Konopka, J. B. (2002). Septin function in Candida albicans morphogenesis. Mol Biol Cell 13, 27322746.
Warenda, A. J., Kauffman, S., Sherrill, T. P., Becker, J. M. & Konopka, J. B. (2003). Candida albicans septin mutants are defective for invasive growth and virulence. Infect Immun 71, 40454051.
Warit, S., Zhang, N., Short, A., Walmsley, R. M., Oliver, S. G. & Stateva, L. I. (2000). Glycosylation deficiency phenotypes resulting from depletion of GDP-mannose pyrophosphorylase in two yeast species. Mol Microbiol 36, 11561166.[CrossRef][Medline]
Zaragoza, O., Rodriguez, C. & Gancedo, C. (2000). Isolation of MIG1 gene from Candida albicans and effects of its disruption on catabolite repression. J Bacteriol 182, 320326.
Ziman, M., Chuang, J. S. & Schekman, R. W. (1996). Chs1p and Chs3p, two proteins involved in chitin synthesis, populate a compartment of the Saccharomyces cerevisiae endocytic pathway. Mol Biol Cell 7, 19091919.[Abstract]
Ziman, M., Chuang, J. S., Tsung, M., Hamamoto, S. & Schekman, R. (1998). Chs6p-dependent anterograde transport of Chs3p from the chitosome to the plasma membrane in Saccharomyces cerevisiae. Mol Biol Cell 9, 15651576.
Received 15 March 2004;
revised 17 May 2004;
accepted 26 May 2004.
HOME | HELP | FEEDBACK | SUBSCRIPTIONS | ARCHIVE | SEARCH | TABLE OF CONTENTS |
INT J SYST EVOL MICROBIOL | MICROBIOLOGY | J GEN VIROL |
J MED MICROBIOL | ALL SGM JOURNALS |