Diversity and evolution of cytochrome P450 monooxygenases in Oomycetes.

Mopeli Marshal Sello,Ntsane Trevor Mthakathi,Mohammad Parvez,Richie Monyaki,Wanping Chen,Jae-Hyuk Yu,Samson Sitheni Mashele,Lehlohonolo Benedict Qhanya,Ipeleng Kopano Rosinah Kgosiemang,Khajamohiddin Syed,Seiso Caiphus Raselemane,David R Nelson,Norventia Jafta

doi:10.1038/srep11572

Abstract

Cytochrome P450 monooxygenases (P450s) are heme-thiolate proteins whose role as drug targets against pathogens, as well as in valuable chemical production and bioremediation, has been explored. In this study we performed comprehensive comparative analysis of P450s in 13 newly explored oomycete pathogens. Three hundred and fifty-six P450s were found in oomycetes. These P450s were grouped into 15 P450 families and 84 P450 subfamilies. Among these, nine P450 families and 31 P450 subfamilies were newly found in oomycetes. Research revealed that oomycetes belonging to different orders contain distinct P450 families and subfamilies in their genomes. Evolutionary analysis and sequence homology data revealed P450 family blooms in oomycetes. Tandem arrangement of a large number of P450s belonging to the same family indicated that P450 family blooming is possibly due to its members’ duplications. A unique combination of amino acid patterns was observed at EXXR and CXG motifs for the P450 families CYP5014, CYP5015 and CYP5017. A novel P450 fusion protein (CYP5619 family) with an N-terminal P450 domain fused to a heme peroxidase/dioxygenase domain was discovered in Saprolegnia declina. Oomycete P450 patterns suggested host influence in shaping their P450 content. This manuscript serves as reference for future P450 annotations in newly explored oomycetes.

Highlights

Cytochrome P450 monooxygenases (P450s) are heme-thiolate proteins whose role as drug targets against pathogens, as well as in valuable chemical production and bioremediation, has been explored
P450s that showed less than 40% identity were assessed for their position on the phylogenetic tree and based on their location/alignment with other P450s they were assigned to different P450 families
Genome-wide identification and annotation of P450s in 13 oomycetes belonging to two different classes and three different orders (Table S1) revealed the presence of a moderate number of P450s in their genomes (Table 1)

Summary

Methods

P450s that showed less than 40% identity were assessed for their position on the phylogenetic tree and based on their location/alignment with other P450s they were assigned to different P450 families. The phylogenetic tree was constructed for evolutionary analysis of oomycete P450s. Identification of P450 family-specific amino acid patterns at EXXR and CXG motifs was carried out using the procedure described elsewhere[41]. After ClustalW alignment of P450s, amino acids in the EXXR and CXG motifs were selected and used for generation of WebLogos and calculation of percentage contribution by an amino acid at each position in the motifs. Annotation and phylogenetic analysis of P450 fused proteins. The selected P450 fused proteins were assigned to different P450 families or subfamilies following the above described criteria. The phylogenetic tree was constructed using MEGA 5.2.242

Results and Discussion

Author Contributions

Additional Information