Abstract
Adenine nucleotide translocase (ANT) is a constitutive mitochondrial component that is involved in ADP/ATP exchange and mitochondrion-mediated apoptosis in yeast and mammals. However, little is known about the function of ANT in pathogenic fungi. In this study, we identified an ANT gene of Puccinia striiformis f. sp. tritici (Pst), designated PsANT. The PsANT protein contains three typical conserved mitochondrion-carrier-protein (mito-carr) domains and shares more than 70% identity with its orthologs from other fungi, suggesting that ANT is conserved in fungi. Immuno-cytochemical localization confirmed the mitochondrial localization of PsANT in normal Pst hyphal cells or collapsed cells. Over-expression of PsANT indicated that PsANT promotes cell death in tobacco, wheat and fission yeast cells. Further study showed that the three mito-carr domains are all needed to induce cell death. qRT-PCR analyses revealed an in-planta induced expression of PsANT during infection. Knockdown of PsANT using a host-induced gene silencing system (HIGS) attenuated the growth and development of virulent Pst at the early infection stage but not enough to alter its pathogenicity. These results provide new insight into the function of PsANT in fungal cell death and growth and might be useful in the search for and design of novel disease control strategies.
Highlights
Cytosolic (c) states, in which ADP-ATP is bound to either the matrix side or the cytosolic side of the inner membrane, respectively
The deduced PsANT protein consisted of 313 amino acids and showed the highest similarity with MlANT (93.9%) in Melampsora larici-populina (Mlp)
A multi-sequence alignment with various adenine nucleotide translocase (ANT) sequences in the National Center of Biotechnology Information database and Broad Institute database revealed that PsANT shared more than 70% identity with its orthologs from other fungi, including Mlp, Puccinia graminis f. sp. tritici (Pgt) and Saccharomyces cerevisiae (Fig. 1a)
Summary
Cytosolic (c) states, in which ADP-ATP is bound to either the matrix side or the cytosolic side of the inner membrane, respectively. The role of ANT in the regulation of cell death by MPT is conserved across nematodes and mammals. Functional analyses of genes in yeasts revealed that the progression of apoptotic –like programmed cell death (PCD) in the single-celled fungus yeast resembles that of higher eukaryotes at the molecular level[14,15]. Comparative analysis of the genome sequence of four Aspergillus and seven other fungal species revealed more than 100 apoptosis-associated genes, including lineage-specific proteins as well as a conserved core component of the ancestral apoptosis machinery that is shared by all fungi, suggesting that there is a complex uncharacterized regulatory network in fungi[21]. Homologs of several mammalian apoptotic proteins, including PARP and AMID, have been found in filamentous fungi[21], indicating that the molecular pathways that control apoptosis in different organisms are conserved even though they differ in complexity. Knockdown of PsANT in Pst using host-induced gene silencing (HIGS) blocked the growth and development of Pst in the early infection stage but did not alter the virulence of Pst
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