2017 |
Garcia-Rico, R O; Gil-Duran, C; Rojas-Aedo, J F; Vaca, I; Figueroa, L; Levican, G; Chavez, R Heterotrimeric G Protein Alpha Subunit Controls Growth, Stress Response, Extracellular Protease Activity, and Cyclopiazonic Acid Production in Penicillium Camemberti Artículo de revista Fungal Biology, 121 (9), pp. 754-762, 2017, ISSN: 1878-6146. Resumen | Enlaces | BibTeX | Etiquetas: alpha-subunit, aspergillus-nidulans, expression, filamentous fungus, g-beta, gene, geotrichum-candidum, growth, metabolism, micotoxin, mycotoxin pathogenicity, pcr, production, proteases, real-time reproduction resistance, secondary stress vegetative @article{RN339, title = {Heterotrimeric G Protein Alpha Subunit Controls Growth, Stress Response, Extracellular Protease Activity, and Cyclopiazonic Acid Production in Penicillium Camemberti}, author = { R.O. Garcia-Rico and C. Gil-Duran and J.F. Rojas-Aedo and I. Vaca and L. Figueroa and G. Levican and R. Chavez}, url = {/brokenurl#<Go to ISI>://WOS:000419930600002}, doi = {10.1016/j.funbio.2017.05.007}, issn = {1878-6146}, year = {2017}, date = {2017-01-01}, journal = {Fungal Biology}, volume = {121}, number = {9}, pages = {754-762}, publisher = {2017 British Mycological Society. Published by Elsevier Ltd.}, abstract = {The fungus Penicillium camemberti is widely used in the ripening of various bloomy-rind cheeses. Several properties of P. camemberti are important in cheese ripening, including conidiation, growth and enzyme production, among others. However, the production of mycotoxins such as cyclopiazonic acid during the ripening process by P. camemberti has raised concerns among consumers that demand food with minimal contamination. Here we show that overexpressing an alpha-subunit from the subgroup I of the heterotrimeric G protein (G alpha i) influences several of these processes: it negatively affects growth in a media dependent manner, triggers conidial germination, reduces the rate of sporulation, affects thermal and osmotic stress resistance, and also extracellular protease and cyclopiazonic acid production. Our results contribute to understanding the biological determinants underlying these biological processes in the economically important fungus P. camemberti.}, keywords = {alpha-subunit, aspergillus-nidulans, expression, filamentous fungus, g-beta, gene, geotrichum-candidum, growth, metabolism, micotoxin, mycotoxin pathogenicity, pcr, production, proteases, real-time reproduction resistance, secondary stress vegetative}, pubstate = {published}, tppubtype = {article} } The fungus Penicillium camemberti is widely used in the ripening of various bloomy-rind cheeses. Several properties of P. camemberti are important in cheese ripening, including conidiation, growth and enzyme production, among others. However, the production of mycotoxins such as cyclopiazonic acid during the ripening process by P. camemberti has raised concerns among consumers that demand food with minimal contamination. Here we show that overexpressing an alpha-subunit from the subgroup I of the heterotrimeric G protein (G alpha i) influences several of these processes: it negatively affects growth in a media dependent manner, triggers conidial germination, reduces the rate of sporulation, affects thermal and osmotic stress resistance, and also extracellular protease and cyclopiazonic acid production. Our results contribute to understanding the biological determinants underlying these biological processes in the economically important fungus P. camemberti. |
2015 |
Gil-Duran, C; Rojas-Aedo, J F; Medina, E; Vaca, I; Garcia-Rico, R O; Villagran, S; Levican, G; Chavez, R The Pcz1 Gene, Which Encodes a Zn(Ii)(2)Cys(6) Protein, Is Involved in the Control of Growth, Conidiation, and Conidial Germination in the Filamentous Fungus Penicillium Roqueforti Artículo de revista Plos One, 10 (3), 2015, ISSN: 1932-6203. Resumen | Enlaces | BibTeX | Etiquetas: alpha-subunit, aspergillus-nidulans, chrysogenum, cluster, pga1 @article{RN240, title = {The Pcz1 Gene, Which Encodes a Zn(Ii)(2)Cys(6) Protein, Is Involved in the Control of Growth, Conidiation, and Conidial Germination in the Filamentous Fungus Penicillium Roqueforti}, author = { C. Gil-Duran and J.F. Rojas-Aedo and E. Medina and I. Vaca and R.O. Garcia-Rico and S. Villagran and G. Levican and R. Chavez}, url = {/brokenurl#<Go to ISI>://WOS:000356353700059}, doi = {10.1371/journal.pone.0120740}, issn = {1932-6203}, year = {2015}, date = {2015-01-01}, journal = {Plos One}, volume = {10}, number = {3}, abstract = {Proteins containing Zn(II)(2)Cys(6) domains are exclusively found in fungi and yeasts. Genes encoding this class of proteins are broadly distributed in fungi, but few of them have been functionally characterized. In this work, we have characterized a gene from the filamentous fungus Penicillium roqueforti that encodes a Zn(II)(2)Cys(6) protein, whose function to date remains unknown. We have named this gene pcz1. We showed that the expression of pcz1 is negatively regulated in a P. roqueforti strain containing a dominant active Gai protein, suggesting that pcz1 encodes a downstream effector that is negatively controlled by Gai. More interestingly, the silencing of pcz1 in P. roqueforti using RNAi-silencing technology resulted in decreased apical growth, the promotion of conidial germination (even in the absence of a carbon source), and the strong repression of conidiation, concomitant with the downregulation of the genes of the central conidiation pathway brlA, abaA and wetA. A model for the participation of pcz1 in these physiological processes in P. roqueforti is proposed.}, keywords = {alpha-subunit, aspergillus-nidulans, chrysogenum, cluster, pga1}, pubstate = {published}, tppubtype = {article} } Proteins containing Zn(II)(2)Cys(6) domains are exclusively found in fungi and yeasts. Genes encoding this class of proteins are broadly distributed in fungi, but few of them have been functionally characterized. In this work, we have characterized a gene from the filamentous fungus Penicillium roqueforti that encodes a Zn(II)(2)Cys(6) protein, whose function to date remains unknown. We have named this gene pcz1. We showed that the expression of pcz1 is negatively regulated in a P. roqueforti strain containing a dominant active Gai protein, suggesting that pcz1 encodes a downstream effector that is negatively controlled by Gai. More interestingly, the silencing of pcz1 in P. roqueforti using RNAi-silencing technology resulted in decreased apical growth, the promotion of conidial germination (even in the absence of a carbon source), and the strong repression of conidiation, concomitant with the downregulation of the genes of the central conidiation pathway brlA, abaA and wetA. A model for the participation of pcz1 in these physiological processes in P. roqueforti is proposed. |
Chavez, R; Fierro, F; Garcia-Rico, R O; Vaca, I Filamentous Fungi from Extreme Environments as a Promising Source of Novel Bioactive Secondary Metabolites Artículo de revista Frontiers in Microbiology, 6 , 2015, ISSN: 1664-302x. Resumen | Enlaces | BibTeX | Etiquetas: aspergillus-nidulans, biosynthetic clusters, discovery, drug environments, expression, extreme filamentous fungi, gene genome heterologous metabolites, metagenome, metagenomics, mining, natural natural-products, polyketide, products, resource, secondary sequence, strategy @article{RN241, title = {Filamentous Fungi from Extreme Environments as a Promising Source of Novel Bioactive Secondary Metabolites}, author = { R. Chavez and F. Fierro and R.O. Garcia-Rico and I. Vaca}, url = {/brokenurl#<Go to ISI>://WOS:000361157000001}, doi = {10.3389/fmicb.2015.00903}, issn = {1664-302x}, year = {2015}, date = {2015-01-01}, journal = {Frontiers in Microbiology}, volume = {6}, abstract = {Natural product search is undergoing resurgence upon the discovery of a huge previously unknown potential for secondary metabolite (SM) production hidden in microbial genomes. This is also the case for filamentous fungi, since their genomes contain a high number of "orphan" SM gene clusters. Recent estimates indicate that only 5% of existing fungal species have been described, thus the potential for the discovery of novel metabolites in fungi is huge. In this context, fungi thriving in harsh environments are of particular interest since they are outstanding producers of unusual chemical structures. At present, there are around 16 genomes from extreme environment-isolated fungi in databases. In a preliminary analysis of three of these genomes we found that several of the predicted SM gene clusters are probably involved in the biosynthesis of compounds not yet described. Genome mining strategies allow the exploitation of the information in genome sequences for the discovery of new natural compounds. The synergy between genome mining strategies and the expected abundance of SMs in fungi from extreme environments is a promising path to discover new natural compounds as a source of medically useful drugs.}, keywords = {aspergillus-nidulans, biosynthetic clusters, discovery, drug environments, expression, extreme filamentous fungi, gene genome heterologous metabolites, metagenome, metagenomics, mining, natural natural-products, polyketide, products, resource, secondary sequence, strategy}, pubstate = {published}, tppubtype = {article} } Natural product search is undergoing resurgence upon the discovery of a huge previously unknown potential for secondary metabolite (SM) production hidden in microbial genomes. This is also the case for filamentous fungi, since their genomes contain a high number of "orphan" SM gene clusters. Recent estimates indicate that only 5% of existing fungal species have been described, thus the potential for the discovery of novel metabolites in fungi is huge. In this context, fungi thriving in harsh environments are of particular interest since they are outstanding producers of unusual chemical structures. At present, there are around 16 genomes from extreme environment-isolated fungi in databases. In a preliminary analysis of three of these genomes we found that several of the predicted SM gene clusters are probably involved in the biosynthesis of compounds not yet described. Genome mining strategies allow the exploitation of the information in genome sequences for the discovery of new natural compounds. The synergy between genome mining strategies and the expected abundance of SMs in fungi from extreme environments is a promising path to discover new natural compounds as a source of medically useful drugs. |
2011 |
Espinosa, Y; Trebotich, J; Sepulveda, F; Cadena, J; Vargas-Straube, M J; Vaca, I; Bull, P; Levican, G; Chavez, R Production of a Heterologous Recombinant Protein Using Fragments of the Glyceraldehyde-3-Phosphate Dehydrogenase Promoter from Penicillium Camemberti Artículo de revista World Journal of Microbiology & Biotechnology, 27 (12), pp. 3019-3023, 2011, ISSN: 0959-3993. Resumen | Enlaces | BibTeX | Etiquetas: aspergillus-nidulans, beta-galactosidase, camemberti, cloning, dehydrogenase elements, expression, functional gene, glyceraldehyde-3-phosphate gpda heterologous nidulans penicillium promoter, protein region sequence-analysis, transformation, yeast @article{RN37c, title = {Production of a Heterologous Recombinant Protein Using Fragments of the Glyceraldehyde-3-Phosphate Dehydrogenase Promoter from Penicillium Camemberti}, author = { Y. Espinosa and J. Trebotich and F. Sepulveda and J. Cadena and M.J. Vargas-Straube and I. Vaca and P. Bull and G. Levican and R. Chavez}, url = {/brokenurl#<Go to ISI>://WOS:000297594400033}, doi = {10.1007/s11274-011-0782-7}, issn = {0959-3993}, year = {2011}, date = {2011-01-01}, journal = {World Journal of Microbiology & Biotechnology}, volume = {27}, number = {12}, pages = {3019-3023}, abstract = {The biotechnological applications of cheese-ripening fungi have been limited by a lack of genetics tools, in particular the identification and characterization of suitable promoters for protein expression. In this study, the suitability of the glyceraldehyde-3-phosphate dehydrogenase (gpdP) promoter from Penicillium camemberti to drive the production of a recombinant protein was evaluated. The gpdP gene and its promoter were isolated using PCR and Genome Walker. The promoter of gpdP has two regions with high identity to the regulatory elements gpd-box and ct-box previously described in Aspergillus nidulans. Two fragments of the promoter containing the gpd- and ct-box or the ct-box alone were used to drive the in vivo production of recombinant beta-galactosidase using A. nidulans as host. Our results indicate that larger fragment containing gpd-box enhances the production of beta-galactosidase activity levels respect to ct-box alone, and that both boxes are necessary to obtain maximal enzymatic activity production. The smaller fragment (187 nt) containing the ct-box alone was able to trigger up to 27% of beta-galactosidase activity, and to our knowledge this is the smallest fragment from a gpd gene used to produce a recombinant protein. Differences were not observed when glycerol, galactose or glucose were used as carbon sources, suggesting that the promoter activity is carbohydrate-independent. This is the first report in which a Penicillium gpd promoter is used for recombinant protein production. Our results open the way for the future development of a system for recombinant proteins expression in the biotechnologically important cheese-ripening fungus P. camemberti.}, keywords = {aspergillus-nidulans, beta-galactosidase, camemberti, cloning, dehydrogenase elements, expression, functional gene, glyceraldehyde-3-phosphate gpda heterologous nidulans penicillium promoter, protein region sequence-analysis, transformation, yeast}, pubstate = {published}, tppubtype = {article} } The biotechnological applications of cheese-ripening fungi have been limited by a lack of genetics tools, in particular the identification and characterization of suitable promoters for protein expression. In this study, the suitability of the glyceraldehyde-3-phosphate dehydrogenase (gpdP) promoter from Penicillium camemberti to drive the production of a recombinant protein was evaluated. The gpdP gene and its promoter were isolated using PCR and Genome Walker. The promoter of gpdP has two regions with high identity to the regulatory elements gpd-box and ct-box previously described in Aspergillus nidulans. Two fragments of the promoter containing the gpd- and ct-box or the ct-box alone were used to drive the in vivo production of recombinant beta-galactosidase using A. nidulans as host. Our results indicate that larger fragment containing gpd-box enhances the production of beta-galactosidase activity levels respect to ct-box alone, and that both boxes are necessary to obtain maximal enzymatic activity production. The smaller fragment (187 nt) containing the ct-box alone was able to trigger up to 27% of beta-galactosidase activity, and to our knowledge this is the smallest fragment from a gpd gene used to produce a recombinant protein. Differences were not observed when glycerol, galactose or glucose were used as carbon sources, suggesting that the promoter activity is carbohydrate-independent. This is the first report in which a Penicillium gpd promoter is used for recombinant protein production. Our results open the way for the future development of a system for recombinant proteins expression in the biotechnologically important cheese-ripening fungus P. camemberti. |
2017 |
Heterotrimeric G Protein Alpha Subunit Controls Growth, Stress Response, Extracellular Protease Activity, and Cyclopiazonic Acid Production in Penicillium Camemberti Artículo de revista Fungal Biology, 121 (9), pp. 754-762, 2017, ISSN: 1878-6146. |
2015 |
The Pcz1 Gene, Which Encodes a Zn(Ii)(2)Cys(6) Protein, Is Involved in the Control of Growth, Conidiation, and Conidial Germination in the Filamentous Fungus Penicillium Roqueforti Artículo de revista Plos One, 10 (3), 2015, ISSN: 1932-6203. |
Filamentous Fungi from Extreme Environments as a Promising Source of Novel Bioactive Secondary Metabolites Artículo de revista Frontiers in Microbiology, 6 , 2015, ISSN: 1664-302x. |
2011 |
Production of a Heterologous Recombinant Protein Using Fragments of the Glyceraldehyde-3-Phosphate Dehydrogenase Promoter from Penicillium Camemberti Artículo de revista World Journal of Microbiology & Biotechnology, 27 (12), pp. 3019-3023, 2011, ISSN: 0959-3993. |