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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 biosynthesis gene cluster and domain structures of pks11 and pks12 in P. marneffei.Each arrow indicates the direction of transcription and relative sizes of the ORFs. Putative function is indicated for each gene. ACP, acyl carrier protein; AT, acyltransferase; KS, ketosynthase; MT, methyltransferase; R, thioester reductase.
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pntd-0001871-g002: Yellow pigment biosynthesis gene cluster and domain structures of pks11 and pks12 in P. marneffei.Each arrow indicates the direction of transcription and relative sizes of the ORFs. Putative function is indicated for each gene. ACP, acyl carrier protein; AT, acyltransferase; KS, ketosynthase; MT, methyltransferase; R, thioester reductase.

Mentions: The pks11 gene is 7780 bp in length. It has one intron of 52 bp (from 607 to 658 bp). The resultant putative mRNA encodes 2575 amino acid residues with a predicted molecular mass of 282.6 kDa. The pks12 gene is 5485 bp in length. It has one intron of 63 bp (from 439 to 501 bp). The resultant putative mRNA encodes 1806 amino acid residues with a predicted molecular mass of 197.9 kDa. PKS11 has one ketosynthase, one acyltransferase, one acyl carrier protein, one methyltransferase and one thioester reductase domains whereas PKS12 only has one ketosynthase, one acyltransferase and one acyl carrier protein domain (Fig. 2). The domain organization of PKS11 and PKS12 showed that both are fungal non-reducing PKSs.


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 biosynthesis gene cluster and domain structures of pks11 and pks12 in P. marneffei.Each arrow indicates the direction of transcription and relative sizes of the ORFs. Putative function is indicated for each gene. ACP, acyl carrier protein; AT, acyltransferase; KS, ketosynthase; MT, methyltransferase; R, thioester reductase.
© Copyright Policy
Related In: Results  -  Collection

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

pntd-0001871-g002: Yellow pigment biosynthesis gene cluster and domain structures of pks11 and pks12 in P. marneffei.Each arrow indicates the direction of transcription and relative sizes of the ORFs. Putative function is indicated for each gene. ACP, acyl carrier protein; AT, acyltransferase; KS, ketosynthase; MT, methyltransferase; R, thioester reductase.
Mentions: The pks11 gene is 7780 bp in length. It has one intron of 52 bp (from 607 to 658 bp). The resultant putative mRNA encodes 2575 amino acid residues with a predicted molecular mass of 282.6 kDa. The pks12 gene is 5485 bp in length. It has one intron of 63 bp (from 439 to 501 bp). The resultant putative mRNA encodes 1806 amino acid residues with a predicted molecular mass of 197.9 kDa. PKS11 has one ketosynthase, one acyltransferase, one acyl carrier protein, one methyltransferase and one thioester reductase domains whereas PKS12 only has one ketosynthase, one acyltransferase and one acyl carrier protein domain (Fig. 2). The domain organization of PKS11 and PKS12 showed that both are fungal non-reducing PKSs.

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