STE11 disruption reveals the central role of a MAPK pathway in dimorphism and mating in Yarrowia lipolytica

Abstract Yarrowia lipolytica is a dimorphic fungus whose morphology is controlled by several factors such as pH and different compounds. To determine if the STE11-mitogen-activated protein kinase (MAPK) pathway plays a role in dimorphism of Y. lipolytica, we isolated the gene encoding a Mapkkk. The...

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Veröffentlicht in:FEMS yeast research 2006-08, Vol.6 (5), p.801-815
Hauptverfasser: Cervantes-Chávez, José A., Ruiz-Herrera, José
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description Abstract Yarrowia lipolytica is a dimorphic fungus whose morphology is controlled by several factors such as pH and different compounds. To determine if the STE11-mitogen-activated protein kinase (MAPK) pathway plays a role in dimorphism of Y. lipolytica, we isolated the gene encoding a Mapkkk. The isolated gene (STE11) has an ORF of 2832 bp without introns, encoding a protein of 944 amino acids, with a theoretical Mr of 100.9 kDa, that exhibits high homology to fungal Mapkkks. Disruption of the STE11 gene was achieved by the pop-in/pop-out procedure. Growth rate and response to osmotic stress or agents affecting wall integrity were unaffected in the deleted mutants, but they lost the capacity to mate and to grow in the mycelial form. Both alterations were reverted by transformation with the wild-type STE11 gene. The Y. lipolytica STE11 gene driven by two different promoters was unable to complement Saccharomyces cerevisiae ste11Δ mutants, although the gene was transcribed. Also, a wild-type MAPKKK gene from Ustilago maydis failed to complement Y. lipolyticaΔste11 mutants. Both negative results were attributed to a failure of the transgenic gene products to interact with the corresponding regulatory and scaffold proteins. This hypothesis was supported by the observation that a truncated version of the U. maydis MAPKKK gene reversed mating and dimorphic defects in the mutants. All these results demonstrate that the MAPK pathway is essential for both morphogenesis and mating in Y. lipolytica.
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To determine if the STE11-mitogen-activated protein kinase (MAPK) pathway plays a role in dimorphism of Y. lipolytica, we isolated the gene encoding a Mapkkk. The isolated gene (STE11) has an ORF of 2832 bp without introns, encoding a protein of 944 amino acids, with a theoretical Mr of 100.9 kDa, that exhibits high homology to fungal Mapkkks. Disruption of the STE11 gene was achieved by the pop-in/pop-out procedure. Growth rate and response to osmotic stress or agents affecting wall integrity were unaffected in the deleted mutants, but they lost the capacity to mate and to grow in the mycelial form. Both alterations were reverted by transformation with the wild-type STE11 gene. The Y. lipolytica STE11 gene driven by two different promoters was unable to complement Saccharomyces cerevisiae ste11Δ mutants, although the gene was transcribed. Also, a wild-type MAPKKK gene from Ustilago maydis failed to complement Y. lipolyticaΔste11 mutants. Both negative results were attributed to a failure of the transgenic gene products to interact with the corresponding regulatory and scaffold proteins. This hypothesis was supported by the observation that a truncated version of the U. maydis MAPKKK gene reversed mating and dimorphic defects in the mutants. All these results demonstrate that the MAPK pathway is essential for both morphogenesis and mating in Y. lipolytica.</description><identifier>ISSN: 1567-1356</identifier><identifier>EISSN: 1567-1364</identifier><identifier>DOI: 10.1111/j.1567-1364.2006.00084.x</identifier><identifier>PMID: 16879430</identifier><language>eng</language><publisher>Oxford, UK: Blackwell Publishing Ltd</publisher><subject>Amino Acid Sequence ; Dimorphism ; Fungi ; Genetic transformation ; Growth rate ; Homology ; Introns ; Kinases ; MAP kinase ; MAP Kinase Kinase Kinases - chemistry ; MAP Kinase Kinase Kinases - genetics ; MAP Kinase Kinase Kinases - physiology ; MAP Kinase Signaling System - physiology ; MAPK pathway ; Mating ; Molecular Sequence Data ; Morphogenesis ; Mutants ; Mycelia ; Open Reading Frames ; Osmotic stress ; Phenotype ; Protein kinase ; Reproduction ; Saccharomyces cerevisiae Proteins ; STE11 gene ; Transcription, Genetic ; Transformation, Genetic ; Ustilago maydis ; Yarrowia - physiology ; Yarrowia lipolytica</subject><ispartof>FEMS yeast research, 2006-08, Vol.6 (5), p.801-815</ispartof><rights>2006 Federation of European Microbiological Societies Published by Blackwell Publishing Ltd. 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To determine if the STE11-mitogen-activated protein kinase (MAPK) pathway plays a role in dimorphism of Y. lipolytica, we isolated the gene encoding a Mapkkk. The isolated gene (STE11) has an ORF of 2832 bp without introns, encoding a protein of 944 amino acids, with a theoretical Mr of 100.9 kDa, that exhibits high homology to fungal Mapkkks. Disruption of the STE11 gene was achieved by the pop-in/pop-out procedure. Growth rate and response to osmotic stress or agents affecting wall integrity were unaffected in the deleted mutants, but they lost the capacity to mate and to grow in the mycelial form. Both alterations were reverted by transformation with the wild-type STE11 gene. The Y. lipolytica STE11 gene driven by two different promoters was unable to complement Saccharomyces cerevisiae ste11Δ mutants, although the gene was transcribed. Also, a wild-type MAPKKK gene from Ustilago maydis failed to complement Y. lipolyticaΔste11 mutants. Both negative results were attributed to a failure of the transgenic gene products to interact with the corresponding regulatory and scaffold proteins. This hypothesis was supported by the observation that a truncated version of the U. maydis MAPKKK gene reversed mating and dimorphic defects in the mutants. All these results demonstrate that the MAPK pathway is essential for both morphogenesis and mating in Y. lipolytica.</description><subject>Amino Acid Sequence</subject><subject>Dimorphism</subject><subject>Fungi</subject><subject>Genetic transformation</subject><subject>Growth rate</subject><subject>Homology</subject><subject>Introns</subject><subject>Kinases</subject><subject>MAP kinase</subject><subject>MAP Kinase Kinase Kinases - chemistry</subject><subject>MAP Kinase Kinase Kinases - genetics</subject><subject>MAP Kinase Kinase Kinases - physiology</subject><subject>MAP Kinase Signaling System - physiology</subject><subject>MAPK pathway</subject><subject>Mating</subject><subject>Molecular Sequence Data</subject><subject>Morphogenesis</subject><subject>Mutants</subject><subject>Mycelia</subject><subject>Open Reading Frames</subject><subject>Osmotic stress</subject><subject>Phenotype</subject><subject>Protein kinase</subject><subject>Reproduction</subject><subject>Saccharomyces cerevisiae Proteins</subject><subject>STE11 gene</subject><subject>Transcription, Genetic</subject><subject>Transformation, Genetic</subject><subject>Ustilago maydis</subject><subject>Yarrowia - physiology</subject><subject>Yarrowia lipolytica</subject><issn>1567-1356</issn><issn>1567-1364</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2006</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><sourceid>ABUWG</sourceid><sourceid>AFKRA</sourceid><sourceid>AZQEC</sourceid><sourceid>BENPR</sourceid><sourceid>CCPQU</sourceid><sourceid>DWQXO</sourceid><sourceid>GNUQQ</sourceid><recordid>eNqNkVtr3DAQhUVoyP0vBEGhb3YlS5a9kJcQcilNSMjlIU9iIs92tbUtR7Kz2X9fObuk0BCoXjQw3zkazSGEcpbyeL7PU56rIuFCyTRjTKWMsVKmrxtk573x5b3O1TbZDWHOGC8it0W2uSqLiRRsh_y-uz_lnFY2-KHrrWupxxeEOtB-htRg23uoqXc1UjelQK-Ob37SDvrZApbUtlHYON_NbGgotBVtoLftr7HxCN67hQVa287Vy94a2Ceb0-iMB-t7jzycnd6fXCSX1-c_To4vEyNFKZNSSCNRZqpQWQV5yQSAVFIZNoFYKgEiw0yxoqwqhYKpYiI440_KCDQY-3vk28q38-55wNDrxgaDdQ0tuiHo-HmW54pH8Os_4NwNvo2z6UyIPBNSFlmkyhVlvAvB41R33jbgl5ozPcah53rctB63rsc49Fsc-jVKD9cPDE8NVn-F6_1H4GgFLGyNy_821mePt7GIcrGSu6H7RJx8nOoPcCWmHw</recordid><startdate>20060801</startdate><enddate>20060801</enddate><creator>Cervantes-Chávez, José A.</creator><creator>Ruiz-Herrera, José</creator><general>Blackwell Publishing Ltd</general><general>Oxford University Press</general><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>3V.</scope><scope>7X7</scope><scope>7XB</scope><scope>88E</scope><scope>8FE</scope><scope>8FH</scope><scope>8FI</scope><scope>8FJ</scope><scope>8FK</scope><scope>ABUWG</scope><scope>AFKRA</scope><scope>AZQEC</scope><scope>BBNVY</scope><scope>BENPR</scope><scope>BHPHI</scope><scope>CCPQU</scope><scope>DWQXO</scope><scope>FYUFA</scope><scope>GHDGH</scope><scope>GNUQQ</scope><scope>HCIFZ</scope><scope>K9.</scope><scope>LK8</scope><scope>M0S</scope><scope>M1P</scope><scope>M7P</scope><scope>PQEST</scope><scope>PQQKQ</scope><scope>PQUKI</scope><scope>PRINS</scope><scope>7X8</scope></search><sort><creationdate>20060801</creationdate><title>STE11 disruption reveals the central role of a MAPK pathway in dimorphism and mating in Yarrowia lipolytica</title><author>Cervantes-Chávez, José A. ; 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To determine if the STE11-mitogen-activated protein kinase (MAPK) pathway plays a role in dimorphism of Y. lipolytica, we isolated the gene encoding a Mapkkk. The isolated gene (STE11) has an ORF of 2832 bp without introns, encoding a protein of 944 amino acids, with a theoretical Mr of 100.9 kDa, that exhibits high homology to fungal Mapkkks. Disruption of the STE11 gene was achieved by the pop-in/pop-out procedure. Growth rate and response to osmotic stress or agents affecting wall integrity were unaffected in the deleted mutants, but they lost the capacity to mate and to grow in the mycelial form. Both alterations were reverted by transformation with the wild-type STE11 gene. The Y. lipolytica STE11 gene driven by two different promoters was unable to complement Saccharomyces cerevisiae ste11Δ mutants, although the gene was transcribed. Also, a wild-type MAPKKK gene from Ustilago maydis failed to complement Y. lipolyticaΔste11 mutants. Both negative results were attributed to a failure of the transgenic gene products to interact with the corresponding regulatory and scaffold proteins. This hypothesis was supported by the observation that a truncated version of the U. maydis MAPKKK gene reversed mating and dimorphic defects in the mutants. All these results demonstrate that the MAPK pathway is essential for both morphogenesis and mating in Y. lipolytica.</abstract><cop>Oxford, UK</cop><pub>Blackwell Publishing Ltd</pub><pmid>16879430</pmid><doi>10.1111/j.1567-1364.2006.00084.x</doi><tpages>15</tpages></addata></record>
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subjects Amino Acid Sequence
Dimorphism
Fungi
Genetic transformation
Growth rate
Homology
Introns
Kinases
MAP kinase
MAP Kinase Kinase Kinases - chemistry
MAP Kinase Kinase Kinases - genetics
MAP Kinase Kinase Kinases - physiology
MAP Kinase Signaling System - physiology
MAPK pathway
Mating
Molecular Sequence Data
Morphogenesis
Mutants
Mycelia
Open Reading Frames
Osmotic stress
Phenotype
Protein kinase
Reproduction
Saccharomyces cerevisiae Proteins
STE11 gene
Transcription, Genetic
Transformation, Genetic
Ustilago maydis
Yarrowia - physiology
Yarrowia lipolytica
title STE11 disruption reveals the central role of a MAPK pathway in dimorphism and mating in Yarrowia lipolytica
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