title: Two Isoforms of Yersinia pestis Plasminogen Activator Pla: Intraspecies Distribution, Intrinsic Disorder Propensity, and Contribution to Virulence.
abstract: It has been shown previously that several endemic Y. pestis isolates with limited virulence contained the I259 isoform of the outer membrane protease Pla, while the epidemic highly virulent strains possessed only the T259 Pla isoform. Our sequence analysis of the pla gene from 118 Y. pestis subsp. microtus strains revealed that the I259 isoform was present exclusively in the endemic strains providing a convictive evidence of more ancestral origin of this isoform. Analysis of the effects of the I259T polymorphism on the intrinsic disorder propensity of Pla revealed that the I259T mutation slightly increases the intrinsic disorder propensity of the C-terminal tail of Pla and makes this protein slightly more prone for disorder-based protein-protein interactions, suggesting that the T259 Pla could be functionally more active than the I259 Pla. This assumption was proven experimentally by assessing the coagulase and fibrinolytic activities of the two Pla isoforms in human plasma, as well as in a direct fluorometric assay with the Pla peptide substrate. The virulence testing of Pla-negative or expressing the I259 and T259 Pla isoforms Y. pestis subsp. microtus and subsp. pestis strains did not reveal any significant difference in LD50 values and dose-dependent survival assays between them by using a subcutaneous route of challenge of mice and guinea pigs or intradermal challenge of mice. However, a significant decrease in time-to-death was observed in animals infected with the epidemic T259 Pla-producing strains as compared to the parent Pla-negative variants. Survival curves of the endemic I259 Pla+ strains fit between them, but significant difference in mean time to death post infection between the Pla-strains and their I259 Pla+ variants could be seen only in the isogenic set of subsp. pestis strains. These findings suggest an essential role for the outer membrane protease Pla evolution in Y. pestis bubonic infection exacerbation that is necessary for intensification of epidemic process from endemic natural focality with sporadic cases in men to rapidly expanding epizootics followed by human epidemic outbreaks, local epidemics or even pandemics.
authors.affiliation: State Research Center for Applied Microbiology and Biotechnology, Obolensk, Moscow Region, Russian Federation.
authors.forename: Svetlana V
authors.initials: SV
authors.lastname: Dentovskaya
authors.affiliation: State Research Center for Applied Microbiology and Biotechnology, Obolensk, Moscow Region, Russian Federation.
authors.forename: Mikhail E
authors.initials: ME
authors.lastname: Platonov
authors.affiliation: State Research Center for Applied Microbiology and Biotechnology, Obolensk, Moscow Region, Russian Federation.
authors.forename: Tat'yana E
authors.initials: TE
authors.lastname: Svetoch
authors.affiliation: State Research Center for Applied Microbiology and Biotechnology, Obolensk, Moscow Region, Russian Federation.
authors.forename: Pavel Kh
authors.initials: PK
authors.lastname: Kopylov
authors.affiliation: State Research Center for Applied Microbiology and Biotechnology, Obolensk, Moscow Region, Russian Federation.
authors.forename: Tat'yana I
authors.initials: TI
authors.lastname: Kombarova
authors.affiliation: State Research Center for Applied Microbiology and Biotechnology, Obolensk, Moscow Region, Russian Federation.
authors.forename: Sergey A
authors.initials: SA
authors.lastname: Ivanov
authors.affiliation: State Research Center for Applied Microbiology and Biotechnology, Obolensk, Moscow Region, Russian Federation.
authors.forename: Rima Z
authors.initials: RZ
authors.lastname: Shaikhutdinova
authors.affiliation: State Research Center for Applied Microbiology and Biotechnology, Obolensk, Moscow Region, Russian Federation.
authors.forename: Lyubov' V
authors.initials: LV
authors.lastname: Kolombet
authors.affiliation: Department of Pathology, University of Texas Medical Branch, Galveston, Texas, United States of America.|Department of Microbiology & Immunology, University of Texas Medical Branch, Galveston, Texas, United States of America.
authors.forename: Sadhana
authors.initials: S
authors.lastname: Chauhan
authors.affiliation: Saint Petersburg Medical Pediatric University, Saint Petersburg, Russia.
authors.forename: Vitaly G
authors.initials: VG
authors.lastname: Ablamunits
authors.affiliation: Department of Pathology, University of Texas Medical Branch, Galveston, Texas, United States of America.|Department of Microbiology & Immunology, University of Texas Medical Branch, Galveston, Texas, United States of America.
authors.forename: Vladimir L
authors.initials: VL
authors.lastname: Motin
authors.affiliation: Department of Molecular Medicine and Byrd Alzheimer's Research Institute, Morsani College of Medicine, University of South Florida, Tampa, Florida, United States of America.|Institute for Biological Instrumentation, Russian Academy of Sciences, Pushchino, Moscow Region, Russian Federation.
authors.forename: Vladimir N
authors.initials: VN
authors.lastname: Uversky
authors.affiliation: State Research Center for Applied Microbiology and Biotechnology, Obolensk, Moscow Region, Russian Federation.
authors.forename: Andrey P
authors.identifier: http://orcid.org/0000-0002-5499-7999
authors.initials: AP
authors.lastname: Anisimov
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001426:Bacterial Proteins / Q000737:chemistry / Q000378:metabolism*; D005260:Female; D006168:Guinea Pigs; D007527:Isoenzymes / Q000378:metabolism*; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D010960:Plasminogen Activators / Q000737:chemistry / Q000378:metabolism*; D017386:Sequence Homology, Amino Acid; D013045:Species Specificity; D014774:Virulence; D015010:Yersinia pestis / Q000201:enzymology* / Q000472:pathogenicity
chemical_list: D001426:Bacterial Proteins; D007527:Isoenzymes; C078140:Pla protease, Yersinia pestis; D010960:Plasminogen Activators