Intracellular Growth Is Dependent on Tyrosine Catabolism in the Dimorphic Fungal Pathogen Penicillium marneffei

Abstract

During infection, pathogens must utilise the available nutrient sources in order to grow while simultaneously evading or tolerating the host’s defence systems. Amino acids are an important nutritional source for pathogenic fungi and can be assimilated from host proteins to provide both carbon and nitrogen. The hpdA gene of the dimorphic fungus Penicillium marneffei, which encodes an enzyme which catalyses the second step of tyrosine catabolism, was identified as up-regulated in pathogenic yeast cells. As well as enabling the fungus to acquire carbon and nitrogen, tyrosine is also a precursor in the formation of two types of protective melanin; DOPA melanin and pyomelanin. Chemical inhibition of HpdA in P. marneffei inhibits ex vivo yeast cell production suggesting that tyrosine is a key nutrient source during infectious growth. The genes required for tyrosine catabolism, including hpdA, are located in a gene cluster and the expression of these genes is induced in the presence of tyrosine. A gene ( hmgR) encoding a Zn(II)2-Cys6 binuclear cluster transcription factor is present within the cluster and is required for tyrosine induced expression and repression in the presence of a preferred nitrogen source. AreA, the GATA-type transcription factor which regulates the global response to limiting nitrogen conditions negatively regulates expression of cluster genes in the absence of tyrosine and is required for nitrogen metabolite repression. Deletion of the tyrosine catabolic genes in the cluster affects growth on tyrosine as either a nitrogen or carbon source and affects pyomelanin, but not DOPA melanin, production. In contrast to other genes of the tyrosine catabolic cluster, deletion of hpdA results in no growth within macrophages. This suggests that the ability to catabolise tyrosine is not required for macrophage infection and that HpdA has an additional novel role to that of tyrosine catabolism and pyomelanin production during growth in host cells.

Author Summary

Fungi that infect humans are a major health problem, especially for those with compromised immune systems. Many fungal infections are extremely difficult to cure and if left untreated are fatal. For successful infection to occur, the fungal pathogen must be able to grow by acquiring and utilising the available nutrient sources within the host whilst evading or tolerating the host’s defence systems. Expression profiling in several pathogenic fungal species has revealed that genes required for tyrosine catabolism are induced specifically in the pathogenic cell type at 37°C. As well as enabling the fungus to acquire carbon and nitrogen intermediates from proteins within the host, tyrosine is also an important precursor in the formation of two different types of melanin, which protects cells against the host’s defence systems. This study shows that the ability to catabolise tyrosine and produce tyrosine derived melanin is not required for the initial stages of fungal infection. However, a novel role for hpdA, which encodes the enzyme which catalyses the second step of tyrosine catabolism, was identified during growth in host cells.