A role for the MAP kinase gene MKC1 in cell wall construction and morphological transitions in Candida albicans
Departamento de Microbiología II, Facultad de Farmacia, Universidad Complutense de Madrid, Plaza de Ramón y Cajal s/n, E-28040 Madrid, Spain Sección Departamental de Microbiología, Facultad de Farmàcia, Universidad de València, Avinguda Vicent Andrés Estellés, E-46100 Burjassot, València, Spain ABST...
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| Veröffentlicht in: | Microbiology (Society for General Microbiology) 1998-02, Vol.144 (2), p.411-424 |
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| creator | Navarro-Garcia, Federico Alonso-Monge, Rebeca Rico, Hortensia Pla, Jesus Sentandreu, Rafael Nombela, Cesar |
| description | Departamento de Microbiología II, Facultad de Farmacia, Universidad Complutense de Madrid, Plaza de Ramón y Cajal s/n, E-28040 Madrid, Spain
Sección Departamental de Microbiología, Facultad de Farmàcia, Universidad de València, Avinguda Vicent Andrés Estellés, E-46100 Burjassot, València, Spain
ABSTRACT
The Candida albicans MKC1 gene encodes a mitogen-activated protein (MAP) kinase, which has been cloned by complementation of the lytic phenotype associated with Saccharomyces cerevisiae slt2 ( mpk1 ) mutants. In this work, the physiological role of this MAP kinase in the pathogenic fungus C. albicans was characterized and a role for MKC1 in the biogenesis of the cell wall suggested based on the following criteria. First, C. albicans mkc1 / mkc1 strains displayed alterations in their cell surfaces under specific conditions as evidenced by scanning electron microscopy. Second, an increase in specific cell wall epitopes ( O -glycosylated mannoprotein) was shown by confocal microscopy in mkc1 / mkc1 mutants. Third, the sensitivity to antifungals which inhibit (1,3)-β-glucan and chitin synthesis was increased in these mutants. In addition, evidence for a role for the MKC1 gene in morphological transitions in C. albicans is presented based on the impairment of pseudohyphal formation of mkc1 / mkc1 strains on Spider medium and on the effect of its overexpression on Sacch. cerevisiae colony morphology on SLADH medium. Using the two-hybrid system, it was also demonstrated that MKC1 is able to interact specifically with Sacch. cerevisiae Mkk1p and Mkk2p, the MAP-kinase kinases of the PKC1 -mediated route of Sacch. cerevisiae , and to activate transcription in Sacch. cerevisiae when bound to a DNA-binding element. These results suggest a role for this MAP kinase in the construction of the cell wall of C. albicans and indicate its potential relevance for the development of novel antifungals.
Author for correspondence: Jesús Pla. Tel: +34 1 3941744. Fax: +34 1 3941745. e-mail: jesuspla@eucmax.sim.ucm.es
Keywords: MAP kinase, pseudohyphal growth, antifungal, two-hybrid system, confocal microscopy |
| doi_str_mv | 10.1099/00221287-144-2-411 |
| format | Article |
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Sección Departamental de Microbiología, Facultad de Farmàcia, Universidad de València, Avinguda Vicent Andrés Estellés, E-46100 Burjassot, València, Spain
ABSTRACT
The Candida albicans MKC1 gene encodes a mitogen-activated protein (MAP) kinase, which has been cloned by complementation of the lytic phenotype associated with Saccharomyces cerevisiae slt2 ( mpk1 ) mutants. In this work, the physiological role of this MAP kinase in the pathogenic fungus C. albicans was characterized and a role for MKC1 in the biogenesis of the cell wall suggested based on the following criteria. First, C. albicans mkc1 / mkc1 strains displayed alterations in their cell surfaces under specific conditions as evidenced by scanning electron microscopy. Second, an increase in specific cell wall epitopes ( O -glycosylated mannoprotein) was shown by confocal microscopy in mkc1 / mkc1 mutants. Third, the sensitivity to antifungals which inhibit (1,3)-β-glucan and chitin synthesis was increased in these mutants. In addition, evidence for a role for the MKC1 gene in morphological transitions in C. albicans is presented based on the impairment of pseudohyphal formation of mkc1 / mkc1 strains on Spider medium and on the effect of its overexpression on Sacch. cerevisiae colony morphology on SLADH medium. Using the two-hybrid system, it was also demonstrated that MKC1 is able to interact specifically with Sacch. cerevisiae Mkk1p and Mkk2p, the MAP-kinase kinases of the PKC1 -mediated route of Sacch. cerevisiae , and to activate transcription in Sacch. cerevisiae when bound to a DNA-binding element. These results suggest a role for this MAP kinase in the construction of the cell wall of C. albicans and indicate its potential relevance for the development of novel antifungals.
Author for correspondence: Jesús Pla. Tel: +34 1 3941744. Fax: +34 1 3941745. e-mail: jesuspla@eucmax.sim.ucm.es
Keywords: MAP kinase, pseudohyphal growth, antifungal, two-hybrid system, confocal microscopy</description><identifier>ISSN: 1350-0872</identifier><identifier>EISSN: 1465-2080</identifier><identifier>DOI: 10.1099/00221287-144-2-411</identifier><identifier>PMID: 9493378</identifier><language>eng</language><publisher>Reading: Soc General Microbiol</publisher><subject>Antifungal Agents - pharmacology ; beta-Galactosidase - metabolism ; Biological and medical sciences ; Calcium-Calmodulin-Dependent Protein Kinases - genetics ; Calcium-Calmodulin-Dependent Protein Kinases - metabolism ; Calcium-Calmodulin-Dependent Protein Kinases - physiology ; Candida albicans ; Candida albicans - enzymology ; Candida albicans - genetics ; Candida albicans - ultrastructure ; Cell Wall - enzymology ; Cell Wall - metabolism ; Cell Wall - ultrastructure ; Chitin - metabolism ; DNA, Fungal - genetics ; Flow Cytometry ; Fluorescent Antibody Technique, Indirect ; Fundamental and applied biological sciences. Psychology ; Fungal Proteins - metabolism ; Gene Expression Regulation, Enzymologic ; Gene Expression Regulation, Fungal ; Glucans - metabolism ; Growth, nutrition, metabolism, transports, enzymes. Molecular biology ; MAP Kinase Kinase 1 ; MAP Kinase Kinase 2 ; Membrane Glycoproteins - metabolism ; Microbiology ; Microscopy, Confocal ; Microscopy, Electron ; Microscopy, Electron, Scanning ; Mitogen-Activated Protein Kinase Kinases ; Mitogen-Activated Protein Kinases ; Mycology ; Plasmids ; Protein Kinase C ; Protein-Serine-Threonine Kinases - genetics ; Protein-Serine-Threonine Kinases - metabolism ; Protein-Tyrosine Kinases - genetics ; Protein-Tyrosine Kinases - metabolism ; Recombination, Genetic ; Saccharomyces cerevisiae - genetics ; Saccharomyces cerevisiae - metabolism ; Transcription, Genetic</subject><ispartof>Microbiology (Society for General Microbiology), 1998-02, Vol.144 (2), p.411-424</ispartof><rights>1998 INIST-CNRS</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c501t-745ae2265e4e19ac45a640cbed01aafff4f87648d4a45c72e302344d3a0a28173</citedby><cites>FETCH-LOGICAL-c501t-745ae2265e4e19ac45a640cbed01aafff4f87648d4a45c72e302344d3a0a28173</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><link.rule.ids>315,781,785,27929,27930</link.rule.ids><backlink>$$Uhttp://pascal-francis.inist.fr/vibad/index.php?action=getRecordDetail&idt=2187979$$DView record in Pascal Francis$$Hfree_for_read</backlink><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/9493378$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Navarro-Garcia, Federico</creatorcontrib><creatorcontrib>Alonso-Monge, Rebeca</creatorcontrib><creatorcontrib>Rico, Hortensia</creatorcontrib><creatorcontrib>Pla, Jesus</creatorcontrib><creatorcontrib>Sentandreu, Rafael</creatorcontrib><creatorcontrib>Nombela, Cesar</creatorcontrib><title>A role for the MAP kinase gene MKC1 in cell wall construction and morphological transitions in Candida albicans</title><title>Microbiology (Society for General Microbiology)</title><addtitle>Microbiology (Reading)</addtitle><description>Departamento de Microbiología II, Facultad de Farmacia, Universidad Complutense de Madrid, Plaza de Ramón y Cajal s/n, E-28040 Madrid, Spain
Sección Departamental de Microbiología, Facultad de Farmàcia, Universidad de València, Avinguda Vicent Andrés Estellés, E-46100 Burjassot, València, Spain
ABSTRACT
The Candida albicans MKC1 gene encodes a mitogen-activated protein (MAP) kinase, which has been cloned by complementation of the lytic phenotype associated with Saccharomyces cerevisiae slt2 ( mpk1 ) mutants. In this work, the physiological role of this MAP kinase in the pathogenic fungus C. albicans was characterized and a role for MKC1 in the biogenesis of the cell wall suggested based on the following criteria. First, C. albicans mkc1 / mkc1 strains displayed alterations in their cell surfaces under specific conditions as evidenced by scanning electron microscopy. Second, an increase in specific cell wall epitopes ( O -glycosylated mannoprotein) was shown by confocal microscopy in mkc1 / mkc1 mutants. Third, the sensitivity to antifungals which inhibit (1,3)-β-glucan and chitin synthesis was increased in these mutants. In addition, evidence for a role for the MKC1 gene in morphological transitions in C. albicans is presented based on the impairment of pseudohyphal formation of mkc1 / mkc1 strains on Spider medium and on the effect of its overexpression on Sacch. cerevisiae colony morphology on SLADH medium. Using the two-hybrid system, it was also demonstrated that MKC1 is able to interact specifically with Sacch. cerevisiae Mkk1p and Mkk2p, the MAP-kinase kinases of the PKC1 -mediated route of Sacch. cerevisiae , and to activate transcription in Sacch. cerevisiae when bound to a DNA-binding element. These results suggest a role for this MAP kinase in the construction of the cell wall of C. albicans and indicate its potential relevance for the development of novel antifungals.
Author for correspondence: Jesús Pla. Tel: +34 1 3941744. Fax: +34 1 3941745. e-mail: jesuspla@eucmax.sim.ucm.es
Keywords: MAP kinase, pseudohyphal growth, antifungal, two-hybrid system, confocal microscopy</description><subject>Antifungal Agents - pharmacology</subject><subject>beta-Galactosidase - metabolism</subject><subject>Biological and medical sciences</subject><subject>Calcium-Calmodulin-Dependent Protein Kinases - genetics</subject><subject>Calcium-Calmodulin-Dependent Protein Kinases - metabolism</subject><subject>Calcium-Calmodulin-Dependent Protein Kinases - physiology</subject><subject>Candida albicans</subject><subject>Candida albicans - enzymology</subject><subject>Candida albicans - genetics</subject><subject>Candida albicans - ultrastructure</subject><subject>Cell Wall - enzymology</subject><subject>Cell Wall - metabolism</subject><subject>Cell Wall - ultrastructure</subject><subject>Chitin - metabolism</subject><subject>DNA, Fungal - genetics</subject><subject>Flow Cytometry</subject><subject>Fluorescent Antibody Technique, Indirect</subject><subject>Fundamental and applied biological sciences. Psychology</subject><subject>Fungal Proteins - metabolism</subject><subject>Gene Expression Regulation, Enzymologic</subject><subject>Gene Expression Regulation, Fungal</subject><subject>Glucans - metabolism</subject><subject>Growth, nutrition, metabolism, transports, enzymes. Molecular biology</subject><subject>MAP Kinase Kinase 1</subject><subject>MAP Kinase Kinase 2</subject><subject>Membrane Glycoproteins - metabolism</subject><subject>Microbiology</subject><subject>Microscopy, Confocal</subject><subject>Microscopy, Electron</subject><subject>Microscopy, Electron, Scanning</subject><subject>Mitogen-Activated Protein Kinase Kinases</subject><subject>Mitogen-Activated Protein Kinases</subject><subject>Mycology</subject><subject>Plasmids</subject><subject>Protein Kinase C</subject><subject>Protein-Serine-Threonine Kinases - genetics</subject><subject>Protein-Serine-Threonine Kinases - metabolism</subject><subject>Protein-Tyrosine Kinases - genetics</subject><subject>Protein-Tyrosine Kinases - metabolism</subject><subject>Recombination, Genetic</subject><subject>Saccharomyces cerevisiae - genetics</subject><subject>Saccharomyces cerevisiae - metabolism</subject><subject>Transcription, Genetic</subject><issn>1350-0872</issn><issn>1465-2080</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>1998</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNqFkcGO0zAQhi0EWnYXXgAJyQeEuARmHCdOjlW1sIhFcICzNXWc1pDYxU614u2ZqmU54oPt0f_NeDy_EC8Q3iL0_TsApVB1pkKtK1VpxEfiEnXbVAo6eMz3uoEKOqOeiqtSfgCwCHghLnrd17XpLkVayZwmL8eU5bLz8vPqq_wZIhUvtz5y_GmNMkTp_DTJe-LNpViWfHBLSFFSHOSc8n6XprQNjia5ZIolHMVyzFszEQaSNG1YjuWZeDLSVPzz83ktvr-_-ba-re6-fPi4Xt1VrgFcKqMb8kq1jdcee3Icthrcxg-AROM46rEzre4GTbpxRvkaVK31UBOQ6tDU1-L1qe4-p18HXxY7h3L8BEWfDsWa3ihe7X9BbLUxTYMMqhPociol-9Huc5gp_7YI9miH_WuHZTussmwHJ708Vz9sZj88pJznz_qrs06Fpzfy8FwoD5jCjhvtGXtzwnZhu7sP2Vs2Zw7cySYkbtj9e_EP6XGfQg</recordid><startdate>19980201</startdate><enddate>19980201</enddate><creator>Navarro-Garcia, Federico</creator><creator>Alonso-Monge, Rebeca</creator><creator>Rico, Hortensia</creator><creator>Pla, Jesus</creator><creator>Sentandreu, Rafael</creator><creator>Nombela, Cesar</creator><general>Soc General Microbiol</general><general>Society for General Microbiology</general><scope>IQODW</scope><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>M7N</scope><scope>7X8</scope></search><sort><creationdate>19980201</creationdate><title>A role for the MAP kinase gene MKC1 in cell wall construction and morphological transitions in Candida albicans</title><author>Navarro-Garcia, Federico ; Alonso-Monge, Rebeca ; Rico, Hortensia ; Pla, Jesus ; Sentandreu, Rafael ; Nombela, Cesar</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c501t-745ae2265e4e19ac45a640cbed01aafff4f87648d4a45c72e302344d3a0a28173</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>1998</creationdate><topic>Antifungal Agents - pharmacology</topic><topic>beta-Galactosidase - metabolism</topic><topic>Biological and medical sciences</topic><topic>Calcium-Calmodulin-Dependent Protein Kinases - genetics</topic><topic>Calcium-Calmodulin-Dependent Protein Kinases - metabolism</topic><topic>Calcium-Calmodulin-Dependent Protein Kinases - physiology</topic><topic>Candida albicans</topic><topic>Candida albicans - enzymology</topic><topic>Candida albicans - genetics</topic><topic>Candida albicans - ultrastructure</topic><topic>Cell Wall - enzymology</topic><topic>Cell Wall - metabolism</topic><topic>Cell Wall - ultrastructure</topic><topic>Chitin - metabolism</topic><topic>DNA, Fungal - genetics</topic><topic>Flow Cytometry</topic><topic>Fluorescent Antibody Technique, Indirect</topic><topic>Fundamental and applied biological sciences. Psychology</topic><topic>Fungal Proteins - metabolism</topic><topic>Gene Expression Regulation, Enzymologic</topic><topic>Gene Expression Regulation, Fungal</topic><topic>Glucans - metabolism</topic><topic>Growth, nutrition, metabolism, transports, enzymes. Molecular biology</topic><topic>MAP Kinase Kinase 1</topic><topic>MAP Kinase Kinase 2</topic><topic>Membrane Glycoproteins - metabolism</topic><topic>Microbiology</topic><topic>Microscopy, Confocal</topic><topic>Microscopy, Electron</topic><topic>Microscopy, Electron, Scanning</topic><topic>Mitogen-Activated Protein Kinase Kinases</topic><topic>Mitogen-Activated Protein Kinases</topic><topic>Mycology</topic><topic>Plasmids</topic><topic>Protein Kinase C</topic><topic>Protein-Serine-Threonine Kinases - genetics</topic><topic>Protein-Serine-Threonine Kinases - metabolism</topic><topic>Protein-Tyrosine Kinases - genetics</topic><topic>Protein-Tyrosine Kinases - metabolism</topic><topic>Recombination, Genetic</topic><topic>Saccharomyces cerevisiae - genetics</topic><topic>Saccharomyces cerevisiae - metabolism</topic><topic>Transcription, Genetic</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Navarro-Garcia, Federico</creatorcontrib><creatorcontrib>Alonso-Monge, Rebeca</creatorcontrib><creatorcontrib>Rico, Hortensia</creatorcontrib><creatorcontrib>Pla, Jesus</creatorcontrib><creatorcontrib>Sentandreu, Rafael</creatorcontrib><creatorcontrib>Nombela, Cesar</creatorcontrib><collection>Pascal-Francis</collection><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>Algology Mycology and Protozoology Abstracts (Microbiology C)</collection><collection>MEDLINE - Academic</collection><jtitle>Microbiology (Society for General Microbiology)</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Navarro-Garcia, Federico</au><au>Alonso-Monge, Rebeca</au><au>Rico, Hortensia</au><au>Pla, Jesus</au><au>Sentandreu, Rafael</au><au>Nombela, Cesar</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>A role for the MAP kinase gene MKC1 in cell wall construction and morphological transitions in Candida albicans</atitle><jtitle>Microbiology (Society for General Microbiology)</jtitle><addtitle>Microbiology (Reading)</addtitle><date>1998-02-01</date><risdate>1998</risdate><volume>144</volume><issue>2</issue><spage>411</spage><epage>424</epage><pages>411-424</pages><issn>1350-0872</issn><eissn>1465-2080</eissn><abstract>Departamento de Microbiología II, Facultad de Farmacia, Universidad Complutense de Madrid, Plaza de Ramón y Cajal s/n, E-28040 Madrid, Spain
Sección Departamental de Microbiología, Facultad de Farmàcia, Universidad de València, Avinguda Vicent Andrés Estellés, E-46100 Burjassot, València, Spain
ABSTRACT
The Candida albicans MKC1 gene encodes a mitogen-activated protein (MAP) kinase, which has been cloned by complementation of the lytic phenotype associated with Saccharomyces cerevisiae slt2 ( mpk1 ) mutants. In this work, the physiological role of this MAP kinase in the pathogenic fungus C. albicans was characterized and a role for MKC1 in the biogenesis of the cell wall suggested based on the following criteria. First, C. albicans mkc1 / mkc1 strains displayed alterations in their cell surfaces under specific conditions as evidenced by scanning electron microscopy. Second, an increase in specific cell wall epitopes ( O -glycosylated mannoprotein) was shown by confocal microscopy in mkc1 / mkc1 mutants. Third, the sensitivity to antifungals which inhibit (1,3)-β-glucan and chitin synthesis was increased in these mutants. In addition, evidence for a role for the MKC1 gene in morphological transitions in C. albicans is presented based on the impairment of pseudohyphal formation of mkc1 / mkc1 strains on Spider medium and on the effect of its overexpression on Sacch. cerevisiae colony morphology on SLADH medium. Using the two-hybrid system, it was also demonstrated that MKC1 is able to interact specifically with Sacch. cerevisiae Mkk1p and Mkk2p, the MAP-kinase kinases of the PKC1 -mediated route of Sacch. cerevisiae , and to activate transcription in Sacch. cerevisiae when bound to a DNA-binding element. These results suggest a role for this MAP kinase in the construction of the cell wall of C. albicans and indicate its potential relevance for the development of novel antifungals.
Author for correspondence: Jesús Pla. Tel: +34 1 3941744. Fax: +34 1 3941745. e-mail: jesuspla@eucmax.sim.ucm.es
Keywords: MAP kinase, pseudohyphal growth, antifungal, two-hybrid system, confocal microscopy</abstract><cop>Reading</cop><pub>Soc General Microbiol</pub><pmid>9493378</pmid><doi>10.1099/00221287-144-2-411</doi><tpages>14</tpages><oa>free_for_read</oa></addata></record> |
| fulltext | fulltext |
| identifier | ISSN: 1350-0872 |
| ispartof | Microbiology (Society for General Microbiology), 1998-02, Vol.144 (2), p.411-424 |
| issn | 1350-0872 1465-2080 |
| language | eng |
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| source | MEDLINE; PubMed Central |
| subjects | Antifungal Agents - pharmacology beta-Galactosidase - metabolism Biological and medical sciences Calcium-Calmodulin-Dependent Protein Kinases - genetics Calcium-Calmodulin-Dependent Protein Kinases - metabolism Calcium-Calmodulin-Dependent Protein Kinases - physiology Candida albicans Candida albicans - enzymology Candida albicans - genetics Candida albicans - ultrastructure Cell Wall - enzymology Cell Wall - metabolism Cell Wall - ultrastructure Chitin - metabolism DNA, Fungal - genetics Flow Cytometry Fluorescent Antibody Technique, Indirect Fundamental and applied biological sciences. Psychology Fungal Proteins - metabolism Gene Expression Regulation, Enzymologic Gene Expression Regulation, Fungal Glucans - metabolism Growth, nutrition, metabolism, transports, enzymes. Molecular biology MAP Kinase Kinase 1 MAP Kinase Kinase 2 Membrane Glycoproteins - metabolism Microbiology Microscopy, Confocal Microscopy, Electron Microscopy, Electron, Scanning Mitogen-Activated Protein Kinase Kinases Mitogen-Activated Protein Kinases Mycology Plasmids Protein Kinase C Protein-Serine-Threonine Kinases - genetics Protein-Serine-Threonine Kinases - metabolism Protein-Tyrosine Kinases - genetics Protein-Tyrosine Kinases - metabolism Recombination, Genetic Saccharomyces cerevisiae - genetics Saccharomyces cerevisiae - metabolism Transcription, Genetic |
| title | A role for the MAP kinase gene MKC1 in cell wall construction and morphological transitions in Candida albicans |
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