2017 |
Torrent, C; Gil-Duran, C; Rojas-Aedo, J F; Medina, E; Vaca, I; Castro, P; Garcia-Rico, R O; Cotoras, M; Mendoza, L; Levican, G; Chavez, R Role of Sfk1 Gene in the Filamentous Fungus Penicillium Roqueforti Artículo de revista Frontiers in Microbiology, 8 , 2017, ISSN: 1664-302x. Resumen | Enlaces | BibTeX | Etiquetas: alpha-subunit, biosynthesis, botrytis-cinerea, changes, chrysogenum, expression, four gene germination, growth kinase, metabolites, of pathways, penicillium phenotypic protein rna-mediated roqueforti, saccharomyces-cerevisiae, secondary signaling silencing, stress, suppressor @article{RN338, title = {Role of Sfk1 Gene in the Filamentous Fungus Penicillium Roqueforti}, author = { C. Torrent and C. Gil-Duran and J.F. Rojas-Aedo and E. Medina and I. Vaca and P. Castro and R.O. Garcia-Rico and M. Cotoras and L. Mendoza and G. Levican and R. Chavez}, url = {/brokenurl#<Go to ISI>://WOS:000417142700001}, doi = {10.3389/fmicb.2017.02424}, issn = {1664-302x}, year = {2017}, date = {2017-01-01}, journal = {Frontiers in Microbiology}, volume = {8}, abstract = {The sfk1 (suppressor of four kinase) gene has been mainly studied in Saccharomyces cerevisiae, where it was shown to be involved in growth and thermal stress resistance. This gene is widely conserved within the phylum Ascomycota. Despite this, to date sfk1 has not been studied in any filamentous fungus. Previously, we found that the orthologous of sfk1 was differentially expressed in a strain of Penicillium roqueforti with an altered phenotype. In this work, we have performed a functional characterization of this gene by using RNAi-silencing technology. The silencing of sfk1 in P. roqueforti resulted in decreased apical growth and the promotion of conidial germination, but interesting, it had no effect on conidiation. In addition, the attenuation of the sfk1 expression sensitized the fungus to osmotic stress, but not to thermal stress. RNA-mediated gene-silencing of sfk1 also affected cell wall integrity in the fungus. Finally, the silencing of sfk1 depleted the production of the main secondary metabolites of P. roqueforti, namely roquefortine C, andrastin A, and mycophenolic acid. To the best of our knowledge this is the first study of the sfk1 gene in filamentous fungi.}, keywords = {alpha-subunit, biosynthesis, botrytis-cinerea, changes, chrysogenum, expression, four gene germination, growth kinase, metabolites, of pathways, penicillium phenotypic protein rna-mediated roqueforti, saccharomyces-cerevisiae, secondary signaling silencing, stress, suppressor}, pubstate = {published}, tppubtype = {article} } The sfk1 (suppressor of four kinase) gene has been mainly studied in Saccharomyces cerevisiae, where it was shown to be involved in growth and thermal stress resistance. This gene is widely conserved within the phylum Ascomycota. Despite this, to date sfk1 has not been studied in any filamentous fungus. Previously, we found that the orthologous of sfk1 was differentially expressed in a strain of Penicillium roqueforti with an altered phenotype. In this work, we have performed a functional characterization of this gene by using RNAi-silencing technology. The silencing of sfk1 in P. roqueforti resulted in decreased apical growth and the promotion of conidial germination, but interesting, it had no effect on conidiation. In addition, the attenuation of the sfk1 expression sensitized the fungus to osmotic stress, but not to thermal stress. RNA-mediated gene-silencing of sfk1 also affected cell wall integrity in the fungus. Finally, the silencing of sfk1 depleted the production of the main secondary metabolites of P. roqueforti, namely roquefortine C, andrastin A, and mycophenolic acid. To the best of our knowledge this is the first study of the sfk1 gene in filamentous fungi. |
2016 |
Del-Cid, A; Gil-Duran, C; Vaca, I; Rojas-Aedo, J F; Garcia-Rico, R O; Levican, G; Chavez, R Identification and Functional Analysis of the Mycophenolic Acid Gene Cluster of Penicillium Roqueforti Artículo de revista Plos One, 11 (1), 2016, ISSN: 1932-6203. Resumen | Enlaces | BibTeX | Etiquetas: biosynthesis, cheese, chrysogenum, dehydrogenase, expression, fungi, immunosuppressant, imp liquid-chromatography, mycotoxins, strains @article{RN290, title = {Identification and Functional Analysis of the Mycophenolic Acid Gene Cluster of Penicillium Roqueforti}, author = { A. Del-Cid and C. Gil-Duran and I. Vaca and J.F. Rojas-Aedo and R.O. Garcia-Rico and G. Levican and R. Chavez}, url = {/brokenurl#<Go to ISI>://WOS:000367888100190}, doi = {10.1371/journal.pone.0147047}, issn = {1932-6203}, year = {2016}, date = {2016-01-01}, journal = {Plos One}, volume = {11}, number = {1}, abstract = {The filamentous fungus Penicillium roqueforti is widely known as the ripening agent of blue-veined cheeses. Additionally, this fungus is able to produce several secondary metabolites, including the meroterpenoid compound mycophenolic acid (MPA). Cheeses ripened with P. roqueforti are usually contaminated with MPA. On the other hand, MPA is a commercially valuable immunosuppressant. However, to date the molecular basis of the production of MPA by P. roqueforti is still unknown. Using a bioinformatic approach, we have identified a genomic region of approximately 24.4 kbp containing a seven-gene cluster that may be involved in the MPA biosynthesis in P. roqueforti. Gene silencing of each of these seven genes (named mpaA, mpaB, mpaC, mpaDE, mpaF, mpaG and mpaH) resulted in dramatic reductions in MPA production, confirming that all of these genes are involved in the biosynthesis of the compound. Interestingly, the mpaF gene, originally described in P. brevicompactum as a MPA self-resistance gene, also exerts the same function in P. roqueforti, suggesting that this gene has a dual function in MPA metabolism. The knowledge of the biosynthetic pathway of MPA in P. roqueforti will be important for the future control of MPA contamination in cheeses and the improvement of MPA production for commercial purposes.}, keywords = {biosynthesis, cheese, chrysogenum, dehydrogenase, expression, fungi, immunosuppressant, imp liquid-chromatography, mycotoxins, strains}, pubstate = {published}, tppubtype = {article} } The filamentous fungus Penicillium roqueforti is widely known as the ripening agent of blue-veined cheeses. Additionally, this fungus is able to produce several secondary metabolites, including the meroterpenoid compound mycophenolic acid (MPA). Cheeses ripened with P. roqueforti are usually contaminated with MPA. On the other hand, MPA is a commercially valuable immunosuppressant. However, to date the molecular basis of the production of MPA by P. roqueforti is still unknown. Using a bioinformatic approach, we have identified a genomic region of approximately 24.4 kbp containing a seven-gene cluster that may be involved in the MPA biosynthesis in P. roqueforti. Gene silencing of each of these seven genes (named mpaA, mpaB, mpaC, mpaDE, mpaF, mpaG and mpaH) resulted in dramatic reductions in MPA production, confirming that all of these genes are involved in the biosynthesis of the compound. Interestingly, the mpaF gene, originally described in P. brevicompactum as a MPA self-resistance gene, also exerts the same function in P. roqueforti, suggesting that this gene has a dual function in MPA metabolism. The knowledge of the biosynthetic pathway of MPA in P. roqueforti will be important for the future control of MPA contamination in cheeses and the improvement of MPA production for commercial purposes. |
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. |
2011 |
Vaca, I; Casqueiro, J; Ullan, R V; Rumbero, A; Chavez, R; Martin, J F Journal of Antibiotics, 64 (6), pp. 447-451, 2011, ISSN: 0021-8820. Resumen | Enlaces | BibTeX | Etiquetas: biosynthesis, cephalosporin-c, chrysogenum, conversion, hplc, isopenicillin liquid-chromatography n, natural-products, penicillium-chrysogenum, purification, stability, storage, system temperature @article{RN36h, title = {A Preparative Method for the Purification of Isopenicillin N from Genetically Blocked Acremonium Chrysogenum Strain Td189: Studies on the Degradation Kinetics and Storage Conditions}, author = { I. Vaca and J. Casqueiro and R.V. Ullan and A. Rumbero and R. Chavez and J.F. Martin}, url = {/brokenurl#<Go to ISI>://WOS:000292090800006}, doi = {10.1038/ja.2011.30}, issn = {0021-8820}, year = {2011}, date = {2011-01-01}, journal = {Journal of Antibiotics}, volume = {64}, number = {6}, pages = {447-451}, abstract = {A protocol for preparative isopenicillin N (IPN) purification, a highly interesting and hitherto unavailable intermediate of the penicillin and cephalosporin biosynthetic pathway due to its high unstability, is described. Culture broths of Acremonium chrysogenum TD189, a strain blocked in cephalosporin biosynthesis that accumulates this metabolite, were treated with acetone and filtered though charcoal and a hydrophobic resin in a single step as tandem columns. The cleared broth was then lyophilized and passed though a Sephadex G-25 column. The last step was the purification to homogeneity of IPN in a semipreparative HPLC equipment and, optionally, a desalting step by Sephadex G-10 column. Once purified, a complete analysis of the stability of the compound and the conditions for its long-term storage was carried out. Our results suggest a first-order model for IPN decomposition for all the pH and temperature analyzed. IPN is more stable at neutral pH, and once lyophilized, can be stored under vacuum and -75 degrees C with a half-life of 770 days. The Journal of Antibiotics (2011); published online 27 April 2011}, keywords = {biosynthesis, cephalosporin-c, chrysogenum, conversion, hplc, isopenicillin liquid-chromatography n, natural-products, penicillium-chrysogenum, purification, stability, storage, system temperature}, pubstate = {published}, tppubtype = {article} } A protocol for preparative isopenicillin N (IPN) purification, a highly interesting and hitherto unavailable intermediate of the penicillin and cephalosporin biosynthetic pathway due to its high unstability, is described. Culture broths of Acremonium chrysogenum TD189, a strain blocked in cephalosporin biosynthesis that accumulates this metabolite, were treated with acetone and filtered though charcoal and a hydrophobic resin in a single step as tandem columns. The cleared broth was then lyophilized and passed though a Sephadex G-25 column. The last step was the purification to homogeneity of IPN in a semipreparative HPLC equipment and, optionally, a desalting step by Sephadex G-10 column. Once purified, a complete analysis of the stability of the compound and the conditions for its long-term storage was carried out. Our results suggest a first-order model for IPN decomposition for all the pH and temperature analyzed. IPN is more stable at neutral pH, and once lyophilized, can be stored under vacuum and -75 degrees C with a half-life of 770 days. The Journal of Antibiotics (2011); published online 27 April 2011 |
2017 |
Role of Sfk1 Gene in the Filamentous Fungus Penicillium Roqueforti Artículo de revista Frontiers in Microbiology, 8 , 2017, ISSN: 1664-302x. |
2016 |
Identification and Functional Analysis of the Mycophenolic Acid Gene Cluster of Penicillium Roqueforti Artículo de revista Plos One, 11 (1), 2016, ISSN: 1932-6203. |
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. |
2011 |
Journal of Antibiotics, 64 (6), pp. 447-451, 2011, ISSN: 0021-8820. |