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Fungi have three tetraspanin families with distinct functions.

Lambou K, Tharreau D, Kohler A, Sirven C, Marguerettaz M, Barbisan C, Sexton AC, Kellner EM, Martin F, Howlett BJ, Orbach MJ, Lebrun MH - BMC Genomics (2008)

Bottom Line: Pls1 is found in ascomycetes and basidiomycetes, whereas Tsp2 is restricted to basidiomycetes and Tsp3 to ascomycetes.A unique copy of each of PLS1 and TSP3 was found in ascomycetes in contrast to TSP2, which has several paralogs in the basidiomycetes, Coprinus cinereus and Laccaria bicolor.Phenotypic analysis of gene replacementmutants Deltatsp3 and Deltatpl1 of M. grisea revealed a reduction of the pathogenicity only on rice, in contrast to Deltapls1 mutants, which are completely non-pathogenic on barley and rice.

View Article: PubMed Central - HTML - PubMed

Affiliation: UMR 5240 CNRS-UCB-INSA-Bayer CropScience, Microbiologie, Adaptation et Pathogénie, Bayer CropScience, 14-20 rue Pierre Baizet, 69263 Lyon Cedex 09, France. karine.lambou@laposte.net

ABSTRACT

Background: Tetraspanins are small membrane proteins that belong to a superfamily encompassing 33 members in human and mouse. These proteins act as organizers of membrane-signalling complexes. So far only two tetraspanin families have been identified in fungi. These are Pls1, which is required for pathogenicity of the plant pathogenic ascomycetes, Magnaporthe grisea, Botrytis cinerea and Colletotrichum lindemuthianum, and Tsp2, whose function is unknown. In this report, we describe a third family of tetraspanins (Tsp3) and a new family of tetraspanin-like proteins (Tpl1) in fungi. We also describe expression of some of these genes in M. grisea and a basidiomycete, Laccaria bicolor, and also their functional analysis in M. grisea.

Results: The exhaustive search for tetraspanins in fungal genomes reveals that higher fungi (basidiomycetes and ascomycetes) contain three families of tetraspanins (Pls1, Tsp2 and Tsp3) with different distribution amongst phyla. Pls1 is found in ascomycetes and basidiomycetes, whereas Tsp2 is restricted to basidiomycetes and Tsp3 to ascomycetes. A unique copy of each of PLS1 and TSP3 was found in ascomycetes in contrast to TSP2, which has several paralogs in the basidiomycetes, Coprinus cinereus and Laccaria bicolor. A tetraspanin-like family (Tpl1) was also identified in ascomycetes. Transcriptional analyses in various tissues of L. bicolor and M. grisea showed that PLS1 and TSP2 are expressed in all tissues in L. bicolor and that TSP3 and TPL1 are overexpressed in the sexual fruiting bodies (perithecia) and mycelia of M. grisea, suggesting that these genes are not pseudogenes. Phenotypic analysis of gene replacementmutants Deltatsp3 and Deltatpl1 of M. grisea revealed a reduction of the pathogenicity only on rice, in contrast to Deltapls1 mutants, which are completely non-pathogenic on barley and rice.

Conclusion: A new tetraspanin family (Tsp3) and a tetraspanin-like protein family (Tpl1) have been identified in fungi. Functional analysis by gene replacement showed that these proteins, as well as Pls1, are involved in the infection process of the plant pathogenic fungus M. grisea. The next challenge will be to decipher the role(s) of tetraspanins in a range of symbiotic, saprophytic and human pathogenic fungi.

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Phylogenetic tree of fungal Pls1, Tsp2 and Tsp3 tetraspanins. The Pls1, Tsp2 and Tsp3 were aligned using ClustalX 1.8. Aligned sequences were analyzed by maximum likelihood using PHYML and RO3G_14268 and RO3G_02583 from R. oryzae as outgroups [see Additional files 4]. Bootstraps values are expressed as percentage of 100 replicates. Mg: Magnaporthe grisea, Cg: Chaetomium globosum, Cl: Colletotrichum lindemuthianum, Nc: Neurospora crassa, Pa: Podospora anserina, Nh: Nectria haematococca, Gz: Gibberella zeae, Fv: Fusarium verticilloides, Tr: Trichoderma reesei, Sn: Stagonospora nodorum, Lm: Leptosphaeria maculans, Cp: Coccidioides posadasii, Bc: Botrytis cinerea, Ss: Sclerotinia sclerotiorum, An: Aspergillus niger, Lb: Laccaria bicolor, Cc: Coprinus cinereus, Pc: Phanerochete chrisosporium, Cn: Cryptococcus neoformans, Ur:Uncinocarpus reesii.
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Figure 5: Phylogenetic tree of fungal Pls1, Tsp2 and Tsp3 tetraspanins. The Pls1, Tsp2 and Tsp3 were aligned using ClustalX 1.8. Aligned sequences were analyzed by maximum likelihood using PHYML and RO3G_14268 and RO3G_02583 from R. oryzae as outgroups [see Additional files 4]. Bootstraps values are expressed as percentage of 100 replicates. Mg: Magnaporthe grisea, Cg: Chaetomium globosum, Cl: Colletotrichum lindemuthianum, Nc: Neurospora crassa, Pa: Podospora anserina, Nh: Nectria haematococca, Gz: Gibberella zeae, Fv: Fusarium verticilloides, Tr: Trichoderma reesei, Sn: Stagonospora nodorum, Lm: Leptosphaeria maculans, Cp: Coccidioides posadasii, Bc: Botrytis cinerea, Ss: Sclerotinia sclerotiorum, An: Aspergillus niger, Lb: Laccaria bicolor, Cc: Coprinus cinereus, Pc: Phanerochete chrisosporium, Cn: Cryptococcus neoformans, Ur:Uncinocarpus reesii.

Mentions: Protein sequences from Pls1, Tsp2 and Tsp3 families were aligned using ClustalX 1.8 [see Additional file 4]. A phylogenetic analysis was then conducted using the PHYML software. The Tpl1 tetraspanin-like family was excluded from this alignment as these proteins are too divergent from Pls1, Tsp2 and Tsp3 proteins to be aligned correctly. The resulting phylogenetic tree (Figure 5) shows that Pls1, Tsp2 and Tsp3 form three distinct families. This phylogenetic tree also shows that the PLS1 genes from ascomycetes and basidiomycetes are orthologs, as the tree is congruent to the corresponding species phylogeny [20]. Similarly, TSP3 genes are orthologs (Figure 5). The TSP2 family consists of numerous paralogs, some being recent as their closest relative gene is in the same species (Tsp2B and Tsp2D from L. bicolor). The Tsp2 cluster is rooted by tetraspanins in the zygomycete, R. oryzae (RO3G_08988/RoTsp2-A and RO3G_17009/RoTsp2-B). The presence of tetraspanins in R. oryzae shows that this family of tetraspanins is ancient and predates the split between zygomycetes and higher fungi.


Fungi have three tetraspanin families with distinct functions.

Lambou K, Tharreau D, Kohler A, Sirven C, Marguerettaz M, Barbisan C, Sexton AC, Kellner EM, Martin F, Howlett BJ, Orbach MJ, Lebrun MH - BMC Genomics (2008)

Phylogenetic tree of fungal Pls1, Tsp2 and Tsp3 tetraspanins. The Pls1, Tsp2 and Tsp3 were aligned using ClustalX 1.8. Aligned sequences were analyzed by maximum likelihood using PHYML and RO3G_14268 and RO3G_02583 from R. oryzae as outgroups [see Additional files 4]. Bootstraps values are expressed as percentage of 100 replicates. Mg: Magnaporthe grisea, Cg: Chaetomium globosum, Cl: Colletotrichum lindemuthianum, Nc: Neurospora crassa, Pa: Podospora anserina, Nh: Nectria haematococca, Gz: Gibberella zeae, Fv: Fusarium verticilloides, Tr: Trichoderma reesei, Sn: Stagonospora nodorum, Lm: Leptosphaeria maculans, Cp: Coccidioides posadasii, Bc: Botrytis cinerea, Ss: Sclerotinia sclerotiorum, An: Aspergillus niger, Lb: Laccaria bicolor, Cc: Coprinus cinereus, Pc: Phanerochete chrisosporium, Cn: Cryptococcus neoformans, Ur:Uncinocarpus reesii.
© Copyright Policy - open-access
Related In: Results  -  Collection

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Show All Figures
getmorefigures.php?uid=PMC2278132&req=5

Figure 5: Phylogenetic tree of fungal Pls1, Tsp2 and Tsp3 tetraspanins. The Pls1, Tsp2 and Tsp3 were aligned using ClustalX 1.8. Aligned sequences were analyzed by maximum likelihood using PHYML and RO3G_14268 and RO3G_02583 from R. oryzae as outgroups [see Additional files 4]. Bootstraps values are expressed as percentage of 100 replicates. Mg: Magnaporthe grisea, Cg: Chaetomium globosum, Cl: Colletotrichum lindemuthianum, Nc: Neurospora crassa, Pa: Podospora anserina, Nh: Nectria haematococca, Gz: Gibberella zeae, Fv: Fusarium verticilloides, Tr: Trichoderma reesei, Sn: Stagonospora nodorum, Lm: Leptosphaeria maculans, Cp: Coccidioides posadasii, Bc: Botrytis cinerea, Ss: Sclerotinia sclerotiorum, An: Aspergillus niger, Lb: Laccaria bicolor, Cc: Coprinus cinereus, Pc: Phanerochete chrisosporium, Cn: Cryptococcus neoformans, Ur:Uncinocarpus reesii.
Mentions: Protein sequences from Pls1, Tsp2 and Tsp3 families were aligned using ClustalX 1.8 [see Additional file 4]. A phylogenetic analysis was then conducted using the PHYML software. The Tpl1 tetraspanin-like family was excluded from this alignment as these proteins are too divergent from Pls1, Tsp2 and Tsp3 proteins to be aligned correctly. The resulting phylogenetic tree (Figure 5) shows that Pls1, Tsp2 and Tsp3 form three distinct families. This phylogenetic tree also shows that the PLS1 genes from ascomycetes and basidiomycetes are orthologs, as the tree is congruent to the corresponding species phylogeny [20]. Similarly, TSP3 genes are orthologs (Figure 5). The TSP2 family consists of numerous paralogs, some being recent as their closest relative gene is in the same species (Tsp2B and Tsp2D from L. bicolor). The Tsp2 cluster is rooted by tetraspanins in the zygomycete, R. oryzae (RO3G_08988/RoTsp2-A and RO3G_17009/RoTsp2-B). The presence of tetraspanins in R. oryzae shows that this family of tetraspanins is ancient and predates the split between zygomycetes and higher fungi.

Bottom Line: Pls1 is found in ascomycetes and basidiomycetes, whereas Tsp2 is restricted to basidiomycetes and Tsp3 to ascomycetes.A unique copy of each of PLS1 and TSP3 was found in ascomycetes in contrast to TSP2, which has several paralogs in the basidiomycetes, Coprinus cinereus and Laccaria bicolor.Phenotypic analysis of gene replacementmutants Deltatsp3 and Deltatpl1 of M. grisea revealed a reduction of the pathogenicity only on rice, in contrast to Deltapls1 mutants, which are completely non-pathogenic on barley and rice.

View Article: PubMed Central - HTML - PubMed

Affiliation: UMR 5240 CNRS-UCB-INSA-Bayer CropScience, Microbiologie, Adaptation et Pathogénie, Bayer CropScience, 14-20 rue Pierre Baizet, 69263 Lyon Cedex 09, France. karine.lambou@laposte.net

ABSTRACT

Background: Tetraspanins are small membrane proteins that belong to a superfamily encompassing 33 members in human and mouse. These proteins act as organizers of membrane-signalling complexes. So far only two tetraspanin families have been identified in fungi. These are Pls1, which is required for pathogenicity of the plant pathogenic ascomycetes, Magnaporthe grisea, Botrytis cinerea and Colletotrichum lindemuthianum, and Tsp2, whose function is unknown. In this report, we describe a third family of tetraspanins (Tsp3) and a new family of tetraspanin-like proteins (Tpl1) in fungi. We also describe expression of some of these genes in M. grisea and a basidiomycete, Laccaria bicolor, and also their functional analysis in M. grisea.

Results: The exhaustive search for tetraspanins in fungal genomes reveals that higher fungi (basidiomycetes and ascomycetes) contain three families of tetraspanins (Pls1, Tsp2 and Tsp3) with different distribution amongst phyla. Pls1 is found in ascomycetes and basidiomycetes, whereas Tsp2 is restricted to basidiomycetes and Tsp3 to ascomycetes. A unique copy of each of PLS1 and TSP3 was found in ascomycetes in contrast to TSP2, which has several paralogs in the basidiomycetes, Coprinus cinereus and Laccaria bicolor. A tetraspanin-like family (Tpl1) was also identified in ascomycetes. Transcriptional analyses in various tissues of L. bicolor and M. grisea showed that PLS1 and TSP2 are expressed in all tissues in L. bicolor and that TSP3 and TPL1 are overexpressed in the sexual fruiting bodies (perithecia) and mycelia of M. grisea, suggesting that these genes are not pseudogenes. Phenotypic analysis of gene replacementmutants Deltatsp3 and Deltatpl1 of M. grisea revealed a reduction of the pathogenicity only on rice, in contrast to Deltapls1 mutants, which are completely non-pathogenic on barley and rice.

Conclusion: A new tetraspanin family (Tsp3) and a tetraspanin-like protein family (Tpl1) have been identified in fungi. Functional analysis by gene replacement showed that these proteins, as well as Pls1, are involved in the infection process of the plant pathogenic fungus M. grisea. The next challenge will be to decipher the role(s) of tetraspanins in a range of symbiotic, saprophytic and human pathogenic fungi.

Show MeSH
Related in: MedlinePlus