Mapping the DNA-Binding Motif of Scabin Toxin, a Guanine Modifying Enzyme from Streptomyces scabies

Scabin is a mono-ADP-ribosyltransferase toxin/enzyme and possible virulence factor produced by the agriculture pathogen, Streptomyces scabies. Recently, molecular dynamic approaches and MD simulations revealed its interaction with both NAD+ and DNA substrates. An Essential Dynamics Analysis identified a crab-claw-like mechanism, including coupled changes in the exposed motifs, and the Rβ1-RLa-NLc-STTβ2-WPN-WARTT-(QxE)ARTT sequence motif was proposed as a catalytic signature of the Pierisin family of DNA-acting toxins. A new fluorescence assay was devised to measure the kinetics for both RNA and DNA substrates. Several protein variants were prepared to probe the Scabin-NAD-DNA molecular model and to reveal the reaction mechanism for the transfer of ADP-ribose to the guanine base in the DNA substrate. The results revealed that there are several lysine and arginine residues in Scabin that are important for binding the DNA substrate; also, key residues such as Asn110 in the mechanism of ADP-ribose transfer to the guanine base were identified. The DNA-binding residues are shared with ScARP from Streptomyces coelicolor but are not conserved with Pierisin-1, suggesting that the modification of guanine bases by ADP-ribosyltransferases is divergent even in the Pierisin family.

Investigation of Streptomyces scabies Causing Potato Scab by Various Detection Techniques, Its Pathogenicity and Determination of Host-Disease Resistance in Potato Germplasm

Streptomyces scabies is a Gram-positive bacterial pathogen that causes common scab disease to several crops, particularly in the potato. It is a soil borne pathogen, a very devastating scab pathogen and difficult to manage in the field. Streptomyces has several species that cause common scab such as S. scabiei, S. acidiscabies, S. europaeiscabiei, S. luridiscabiei, S. niveiscabiei, S. puniciscabiei, S. reticuliscabiei, S. stelliscabiei, S. turgidiscabies, S. ipomoeae. Common scab disease harmfully affects potato economic and market value due to the presence of black spots on the tuber. Owing to its genetic diversity and pathogenicity, the determination of pathogen presence in potato fields is still challenging. In this study, S. scabies genetic diversity was measured by surveying five potato-growing areas of Pakistan during the growing season 2019. A total of 50 Streptomyces isolates, including S. scabies, S. acidiscabies, S. griseoflavus were isolated and identified based on morphologic, biochemical and molecular analysis. Virulent confirmation assays confirmed ten virulent strains of Streptomyces spp. On the potato cultivars Cardinal and Santee. Among the Streptomyces species, S. scabies showed the highest scab index, followed by S. acidiscabies and S. griseoflavus by exhibiting the scab-like lesions on potato tubers. Ten potato cultivars were screened against these virulent isolates of Streptomyces. The Faisalabad white variety showed the highest scab index followed By Cardinal, Tourag, Kuroda, Santee, Lady Rosetta, Asterix, Diamant, Faisalabad red and Sadaf. Moreover, genetic diversity and pathogenicity of Streptomyces spp. on potato tubers were also likely diverse in different geographical regions and also potato cultivars. This study represents a contribution to understanding the local interaction between potatoes and Streptomyces spp. in Pakistan. It will aid in supporting a solution for the management of this pathogen around the world.