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First discovery of two polyketide synthase genes for mitorubrinic acid and mitorubrinol yellow pigment biosynthesis and implications in virulence of Penicillium marneffei.

Woo PC, Lam CW, Tam EW, Leung CK, Wong SS, Lau SK, Yuen KY - PLoS Negl Trop Dis (2012)

Bottom Line: Sequence analysis showed that PKS11 and PKS12 are fungal non-reducing PKSs.The survival of mice challenged with the pks11 knockdown, pks12 knockdown and pks11pks12 double knockdown mutants was significantly better than those challenged with wild type P. marneffei (P<0.05).There was also statistically significant decrease in survival of pks11 knockdown, pks12 knockdown and pks11pks12 double knockdown mutants compared to wild type P. marneffei in both J774 and THP1 macrophages (P<0.05).

View Article: PubMed Central - PubMed

Affiliation: State Key Laboratory of Emerging Infectious Diseases, The University of Hong Kong, Hong Kong.

ABSTRACT

Background: The genome of P. marneffei, the most important thermal dimorphic fungus causing respiratory, skin and systemic mycosis in China and Southeast Asia, possesses 23 polyketide synthase (PKS) genes and 2 polyketide synthase nonribosomal peptide synthase hybrid (PKS-NRPS) genes, which is of high diversity compared to other thermal dimorphic pathogenic fungi. We hypothesized that the yellow pigment in the mold form of P. marneffei could also be synthesized by one or more PKS genes.

Methodology/principal findings: All 23 PKS and 2 PKS-NRPS genes of P. marneffei were systematically knocked down. A loss of the yellow pigment was observed in the mold form of the pks11 knockdown, pks12 knockdown and pks11pks12 double knockdown mutants. Sequence analysis showed that PKS11 and PKS12 are fungal non-reducing PKSs. Ultra high performance liquid chromatography-photodiode array detector/electrospray ionization-quadruple time of flight-mass spectrometry (MS) and MS/MS analysis of the culture filtrates of wild type P. marneffei and the pks11 knockdown, pks12 knockdown and pks11pks12 double knockdown mutants showed that the yellow pigment is composed of mitorubrinic acid and mitorubrinol. The survival of mice challenged with the pks11 knockdown, pks12 knockdown and pks11pks12 double knockdown mutants was significantly better than those challenged with wild type P. marneffei (P<0.05). There was also statistically significant decrease in survival of pks11 knockdown, pks12 knockdown and pks11pks12 double knockdown mutants compared to wild type P. marneffei in both J774 and THP1 macrophages (P<0.05).

Conclusions/significance: The yellow pigment of the mold form of P. marneffei is composed of mitorubrinol and mitorubrinic acid. This represents the first discovery of PKS genes responsible for mitorubrinol and mitorubrinic acid biosynthesis. pks12 and pks11 are probably responsible for sequential use in the biosynthesis of mitorubrinol and mitorubrinic acid. Mitorubrinol and mitorubrinic acid are virulence factors of P. marneffei by improving its intracellular survival in macrophages.

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Related in: MedlinePlus

Yellow pigment production of wild type, pks11, pks12 and pks11pks12 knockdown mutants of P. marneffei.(A) Wild type, (B) pks11, (C) pks12 and (D) pks11pks12 knockdown mutants of P. marneffei were grown on Sabouraud dextrose agar after 7 days incubation at 25°C.
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pntd-0001871-g001: Yellow pigment production of wild type, pks11, pks12 and pks11pks12 knockdown mutants of P. marneffei.(A) Wild type, (B) pks11, (C) pks12 and (D) pks11pks12 knockdown mutants of P. marneffei were grown on Sabouraud dextrose agar after 7 days incubation at 25°C.

Mentions: All 23 PKS and 2 PKS-NRPS genes of P. marneffei were systematically knocked down. The median transcription levels of the 25 knockdown mutants were 12.6% (range 0.1% to 40%) of that in wild type. A loss of the yellow pigment was observed exclusively in the mold form of the pks11 and pks12 knockdown mutants, which have pks11 and pks12 transcription levels 5.4% and 10.0% respectively of that in wild type (Fig. 1). A pks11pks12 double knockdown mutant was also constructed. A loss of the yellow pigment was also observed (Fig. 1). The transcription levels of pks11 and pks12 were 26.5% and 9.0% respectively of that in wild type.


First discovery of two polyketide synthase genes for mitorubrinic acid and mitorubrinol yellow pigment biosynthesis and implications in virulence of Penicillium marneffei.

Woo PC, Lam CW, Tam EW, Leung CK, Wong SS, Lau SK, Yuen KY - PLoS Negl Trop Dis (2012)

Yellow pigment production of wild type, pks11, pks12 and pks11pks12 knockdown mutants of P. marneffei.(A) Wild type, (B) pks11, (C) pks12 and (D) pks11pks12 knockdown mutants of P. marneffei were grown on Sabouraud dextrose agar after 7 days incubation at 25°C.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC3475676&req=5

pntd-0001871-g001: Yellow pigment production of wild type, pks11, pks12 and pks11pks12 knockdown mutants of P. marneffei.(A) Wild type, (B) pks11, (C) pks12 and (D) pks11pks12 knockdown mutants of P. marneffei were grown on Sabouraud dextrose agar after 7 days incubation at 25°C.
Mentions: All 23 PKS and 2 PKS-NRPS genes of P. marneffei were systematically knocked down. The median transcription levels of the 25 knockdown mutants were 12.6% (range 0.1% to 40%) of that in wild type. A loss of the yellow pigment was observed exclusively in the mold form of the pks11 and pks12 knockdown mutants, which have pks11 and pks12 transcription levels 5.4% and 10.0% respectively of that in wild type (Fig. 1). A pks11pks12 double knockdown mutant was also constructed. A loss of the yellow pigment was also observed (Fig. 1). The transcription levels of pks11 and pks12 were 26.5% and 9.0% respectively of that in wild type.

Bottom Line: Sequence analysis showed that PKS11 and PKS12 are fungal non-reducing PKSs.The survival of mice challenged with the pks11 knockdown, pks12 knockdown and pks11pks12 double knockdown mutants was significantly better than those challenged with wild type P. marneffei (P<0.05).There was also statistically significant decrease in survival of pks11 knockdown, pks12 knockdown and pks11pks12 double knockdown mutants compared to wild type P. marneffei in both J774 and THP1 macrophages (P<0.05).

View Article: PubMed Central - PubMed

Affiliation: State Key Laboratory of Emerging Infectious Diseases, The University of Hong Kong, Hong Kong.

ABSTRACT

Background: The genome of P. marneffei, the most important thermal dimorphic fungus causing respiratory, skin and systemic mycosis in China and Southeast Asia, possesses 23 polyketide synthase (PKS) genes and 2 polyketide synthase nonribosomal peptide synthase hybrid (PKS-NRPS) genes, which is of high diversity compared to other thermal dimorphic pathogenic fungi. We hypothesized that the yellow pigment in the mold form of P. marneffei could also be synthesized by one or more PKS genes.

Methodology/principal findings: All 23 PKS and 2 PKS-NRPS genes of P. marneffei were systematically knocked down. A loss of the yellow pigment was observed in the mold form of the pks11 knockdown, pks12 knockdown and pks11pks12 double knockdown mutants. Sequence analysis showed that PKS11 and PKS12 are fungal non-reducing PKSs. Ultra high performance liquid chromatography-photodiode array detector/electrospray ionization-quadruple time of flight-mass spectrometry (MS) and MS/MS analysis of the culture filtrates of wild type P. marneffei and the pks11 knockdown, pks12 knockdown and pks11pks12 double knockdown mutants showed that the yellow pigment is composed of mitorubrinic acid and mitorubrinol. The survival of mice challenged with the pks11 knockdown, pks12 knockdown and pks11pks12 double knockdown mutants was significantly better than those challenged with wild type P. marneffei (P<0.05). There was also statistically significant decrease in survival of pks11 knockdown, pks12 knockdown and pks11pks12 double knockdown mutants compared to wild type P. marneffei in both J774 and THP1 macrophages (P<0.05).

Conclusions/significance: The yellow pigment of the mold form of P. marneffei is composed of mitorubrinol and mitorubrinic acid. This represents the first discovery of PKS genes responsible for mitorubrinol and mitorubrinic acid biosynthesis. pks12 and pks11 are probably responsible for sequential use in the biosynthesis of mitorubrinol and mitorubrinic acid. Mitorubrinol and mitorubrinic acid are virulence factors of P. marneffei by improving its intracellular survival in macrophages.

Show MeSH
Related in: MedlinePlus