Shaikhutdinova RZ, Ivanov SA, Dentovskaya SV, Titareva GM, Knirel YA Characterization of a transposon Tn5-generated mutant of Yersinia pestis defective in lipooligosaccharide biosynthesis Biochemistry (Moscow)84(4) (2019)
398-407
The structure was elucidated in this paper NCBI PubMed ID:31228931 Publication DOI:10.1134/S0006297919040072 Journal NLM ID:0376536 Publisher: Nauka/Interperiodica Correspondence: S. V. Dentovskaya <dentovskayaobolensk.org> Institutions: Zelinsky Institute of Organic Chemistry, Russian Academy of Sciences, Moscow, Russia, State Research Center for Applied Microbiology and Biotechnology, Obolensk, Moscow Region, 142279, Russia
To identify Yersinia pestis genes involved in the microbe's resistance to cationic antimicrobial peptides, the strategy of random transposon mutagenesis with a Tn5 minitransposon was used, and the library was screened for detecting polymyxin B (PMB) susceptible mutants. The mutation responsible for PMB-sensitive phenotype and the lipopolysaccharide (LPS) structure were characterized for the Y. pestis strain KM218-A3. In this strain the mini-Tn5 was located in an open reading frame with the product homologous to the E. coli protein GmhB (82% identity) functioning as d-glycero-d-manno-heptose-1,7-diphosphate phosphatase. ESI FT ICR mass spectrometry of anions was used to study the structure of the unmodified LPS of Y. pestis KM218-A3, and molecules were revealed with the full-size LPS core or with two types of an incomplete core: consisting of Kdo-Kdo or Ko-Kdo disaccharides and Hep-(Kdo)-Kdo or Hep-(Ko)-Kdo trisaccharides. The performed complementation confirmed that the defect in the biological properties of the mutant strain was caused by inactivation of the gmhB gene. These findings indicated that the gmhB gene product of Y. pestis is essential for production of wild-type LPS resistant to antimicrobial peptides and serum.
Methods: DNA sequencing, SDS-PAGE, DNA techniques, ESI-ICR-MS, serological methods, insertion mutagenesis, complementation Comments, role: Yersinia pestis mutant strain KM218-A3; minor glycoform
Related record ID(s): 2281, 2282, 2283, 2284, 2285, 2286, 2287, 2288, 2289, 2290 NCBI Taxonomy refs (TaxIDs):632 Reference(s) to other database(s): GTC:G76532QK Show glycosyltransferases
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