title: Contributions of environmental signals and conserved residues to the functions of carbon storage regulator A of Borrelia burgdorferi.
abstract: Carbon storage regulator A of Borrelia burgdorferi (CsrABb) contributes to vertebrate host-specific adaptation by modulating activation of the Rrp2-RpoN-RpoS pathway and is critical for infectivity. We hypothesized that the functions of CsrABb are dependent on environmental signals and on select residues. We analyzed the phenotype of csrABb deletion and site-specific mutants to determine the conserved and pathogen-specific attributes of CsrABb. Levels of phosphate acetyltransferase (Pta) involved in conversion of acetyl phosphate to acetyl-coenzyme A (acetyl-CoA) and posttranscriptionally regulated by CsrABb in the csrABb mutant were reduced from or similar to those in the control strains under unfed- or fed-tick conditions, respectively. Increased levels of supplemental acetate restored vertebrate host-responsive determinants in the csrABb mutant to parental levels, indicating that both the levels of CsrABb and the acetyl phosphate and acetyl-CoA balance contribute to the activation of the Rrp2-RpoN-RpoS pathway. Site-specific replacement of 8 key residues of CsrABb (8S) with alanines resulted in increased levels of CsrABb and reduced levels of Pta and acetyl-CoA, while levels of RpoS, BosR, and other members of rpoS regulon were elevated. Truncation of 7 amino acids at the C terminus of CsrABb (7D) resulted in reduced csrABb transcripts and posttranscriptionally reduced levels of FliW located upstream of CsrABb. Electrophoretic mobility shift assays revealed increased binding of 8S mutant protein to the CsrA binding box upstream of pta compared to the parental and 7D truncated protein. Two CsrABb binding sites were also identified upstream of fliW within the flgK coding sequence. These observations reveal conserved and unique functions of CsrABb that regulate adaptive gene expression in B. burgdorferi.
authors.affiliation: South Texas Center for Emerging Infectious Diseases, Center of Excellence in Infection Genomics and Department of Biology, The University of Texas at San Antonio, San Antonio, Texas, USA.
authors.forename: S L Rajasekhar
authors.initials: SL
authors.lastname: Karna
authors.forename: Rajesh G
authors.initials: RG
authors.lastname: Prabhu
authors.forename: Ying-Han
authors.initials: YH
authors.lastname: Lin
authors.forename: Christine L
authors.initials: CL
authors.lastname: Miller
authors.forename: J
authors.initials: J
authors.lastname: Seshu
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism*; D001483:Base Sequence; D015153:Blotting, Western; D025065:Borrelia burgdorferi / Q000235:genetics / Q000378:metabolism* / Q000472:pathogenicity*; D017124:Conserved Sequence; D024202:Electrophoretic Mobility Shift Assay; D015964:Gene Expression Regulation, Bacterial / Q000502:physiology; D006790:Host-Parasite Interactions / Q000502:physiology*; D008969:Molecular Sequence Data; D060888:Real-Time Polymerase Chain Reaction; D013987:Ticks / Q000382:microbiology
chemical_list: D001426:Bacterial Proteins